Interspersed Repetitive Sequences
Copies of transposable elements interspersed throughout the genome, some of which are still active and often referred to as "jumping genes". There are two classes of interspersed repetitive elements. Class I elements (or RETROELEMENTS - such as retrotransposons, retroviruses, LONG INTERSPERSED NUCLEOTIDE ELEMENTS and SHORT INTERSPERSED NUCLEOTIDE ELEMENTS) transpose via reverse transcription of an RNA intermediate. Class II elements (or DNA TRANSPOSABLE ELEMENTS - such as transposons, Tn elements, insertion sequence elements and mobile gene cassettes of bacterial integrons) transpose directly from one site in the DNA to another.
Any of the covalently closed DNA molecules found in bacteria, many viruses, mitochondria, plastids, and plasmids. Small, polydisperse circular DNA's have also been observed in a number of eukaryotic organisms and are suggested to have homology with chromosomal DNA and the capacity to be inserted into, and excised from, chromosomal DNA. It is a fragment of DNA formed by a process of looping out and deletion, containing a constant region of the mu heavy chain and the 3'-part of the mu switch region. Circular DNA is a normal product of rearrangement among gene segments encoding the variable regions of immunoglobulin light and heavy chains, as well as the T-cell receptor. (Riger et al., Glossary of Genetics, 5th ed & Segen, Dictionary of Modern Medicine, 1992)
Molecular Sequence Data
Descriptions of specific amino acid, carbohydrate, or nucleotide sequences which have appeared in the published literature and/or are deposited in and maintained by databanks such as GENBANK, European Molecular Biology Laboratory (EMBL), National Biomedical Research Foundation (NBRF), or other sequence repositories.
Encyclopedias as Topic
Drug Resistance, Microbial
A parasexual process in BACTERIA; ALGAE; FUNGI; and ciliate EUKARYOTA for achieving exchange of chromosome material during fusion of two cells. In bacteria, this is a uni-directional transfer of genetic material; in protozoa it is a bi-directional exchange. In algae and fungi, it is a form of sexual reproduction, with the union of male and female gametes.
Drug Resistance, Bacterial
Messages between computer users via COMPUTER COMMUNICATION NETWORKS. This feature duplicates most of the features of paper mail, such as forwarding, multiple copies, and attachments of images and other file types, but with a speed advantage. The term also refers to an individual message sent in this way.
Biological Science Disciplines
All of the divisions of the natural sciences dealing with the various aspects of the phenomena of life and vital processes. The concept includes anatomy and physiology, biochemistry and biophysics, and the biology of animals, plants, and microorganisms. It should be differentiated from BIOLOGY, one of its subdivisions, concerned specifically with the origin and life processes of living organisms.
Semiautonomous, self-reproducing organelles that occur in the cytoplasm of all cells of most, but not all, eukaryotes. Each mitochondrion is surrounded by a double limiting membrane. The inner membrane is highly invaginated, and its projections are called cristae. Mitochondria are the sites of the reactions of oxidative phosphorylation, which result in the formation of ATP. They contain distinctive RIBOSOMES, transfer RNAs (RNA, TRANSFER); AMINO ACYL T RNA SYNTHETASES; and elongation and termination factors. Mitochondria depend upon genes within the nucleus of the cells in which they reside for many essential messenger RNAs (RNA, MESSENGER). Mitochondria are believed to have arisen from aerobic bacteria that established a symbiotic relationship with primitive protoeukaryotes. (King & Stansfield, A Dictionary of Genetics, 4th ed)
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Journal Impact Factor
Stabilization of poly-L-lysine/DNA polyplexes for in vivo gene delivery to the liver. (1/43669)We are developing a self-assembling non-viral in vivo gene delivery vehicle based on poly-l-lysine and plasmid DNA. We have characterized poly-l-lysines of different chain lengths for DNA condensation and strength of DNA binding. Poly-l-lysine chains >20 residues bound DNA efficiently in physiological saline, while shorter chains did not. Attachment of asialoorosomucoid to PLL increased the PLL chain length required for efficient DNA binding in saline and for efficient DNA condensation. By electron microscopy, poly-l-lysine/DNA polyplexes appeared as toroids 25-50 nm in diameter or rods 40-80 nm long; conjugation of asialoorosomucoid to the polylysine component increased the size of resulting polyplexes to 50-90 nm. In water, poly-l-lysine and asialoorosomucoid-PLL polyplexes have effective diameters of 46 and 87.6 nm, respectively. Polyplexes containing only poly-l-lysine and DNA aggregated in physiological saline at all charge ratios and aggregated at neutral charge ratios in water. Attachment of asialoorosomucoid lessened, but did not eliminate, the aggregation of PLL polyplexes, and did not result in efficient delivery of polyplexes to hepatocytes. Conjugation of polyethylene glycol to poly-l-lysine sterically stabilized resulting polyplexes at neutral charge ratios by shielding the surfaces. For efficient in vivo gene delivery, polyplexes will need to be sterically stabilized to prevent aggregation and interaction with serum components. (+info)
Cloning and characterisation of a novel ompB operon from Vibrio cholerae 569B. (2/43669)The ompB operon of Vibrio cholerae 569B has been cloned and fully sequenced. The operon encodes two proteins, OmpR and EnvZ, which share sequence identity with the OmpR and EnvZ proteins of a variety of other bacteria. Although the order of the ompR and envZ genes of V. cholerae is similar to that of the ompB operon of E. coli, S. typhimurium and X. nematophilus, the Vibrio operon exhibits a number of novel features. The structural organisation and features of the V. cholerae ompB operon are described. (+info)
Co-expression of glutathione S-transferase with methionine aminopeptidase: a system of producing enriched N-terminal processed proteins in Escherichia coli. (3/43669)We describe here an Escherichia coli expression system that produces recombinant proteins enriched in the N-terminal processed form, by using glutathione S-transferase cGSTM1-1 and rGSTT1-1 as models, where c and r refer to chick and rat respectively. Approximately 90% of the cGSTM1-1 or rGSTT1-1 overexpressed in E. coli under the control of a phoA promoter retained the initiator methionine residue that was absent from the mature isoenzymes isolated from tissues. The amount of initiator methionine was decreased to 40% of the expressed cGSTM1-1 when the isoenzyme was co-expressed with an exogenous methionine aminopeptidase gene under the control of a separate phoA promoter. The recombinant proteins expressed were mainly methionine aminopeptidase. The yield of cGSTM1-1 was decreased to 10% of that expressed in the absence of the exogenous methionine aminopeptidase gene. By replacing the phoA with its natural promoter, the expression of methionine aminopeptidase decreased drastically. The yield of the co-expressed cGSTM1-1 was approx. 60% of that in the absence of the exogenous methionine aminopeptidase gene; approx. 65% of the initiator methionine residues were removed from the enzyme. Under similar conditions, N-terminal processing was observed in approx. 70% of the recombinant rGSTT1-1 expressed. By increasing the concentration of phosphate in the growth medium, the amount of initiator methionine on cGSTM1-1 was decreased to 14% of the overexpressed isoenzymes, whereas no further improvement could be observed for rGSTT1-1. The initiator methionine residue does not affect the enzymic activities of either cGSTM1-1 or rGSTT1-1. However, the epoxidase activity and the 4-nitrobenzyl chloride-conjugating activity of the purified recombinant rGSTT1-1 are markedly higher that those reported recently for the same isoenzyme isolated from rat livers. (+info)
Expression of the plague plasminogen activator in Yersinia pseudotuberculosis and Escherichia coli. (4/43669)Enteropathogenic yersiniae (Yersinia pseudotuberculosis and Yersinia enterocolitica) typically cause chronic disease as opposed to the closely related Yersinia pestis, the causative agent of bubonic plague. It is established that this difference reflects, in part, carriage by Y. pestis of a unique 9.6-kb pesticin or Pst plasmid (pPCP) encoding plasminogen activator (Pla) rather than distinctions between shared approximately 70-kb low-calcium-response, or Lcr, plasmids (pCD in Y. pestis and pYV in enteropathogenic yersiniae) encoding cytotoxic Yops and anti-inflammatory V antigen. Pla is known to exist as a combination of 32.6-kDa (alpha-Pla) and slightly smaller (beta-Pla) outer membrane proteins, of which at least one promotes bacterial dissemination in vivo and degradation of Yops in vitro. We show here that only alpha-Pla accumulates in Escherichia coli LE392/pPCP1 cultivated in enriched medium and that either autolysis or extraction of this isolate with 1.0 M NaCl results in release of soluble alpha and beta forms possessing biological activity. This process also converted cell-bound alpha-Pla to beta-Pla and smaller forms in Y. pestis KIM/pPCP1 and Y. pseudotuberculosis PB1/+/pPCP1 but did not promote solubilization. Pla-mediated posttranslational hydrolysis of pulse-labeled Yops in Y. pseudotuberculosis PB1/+/pPCP1 occurred more slowly than that in Y. pestis but was otherwise similar except for accumulation of stable degradation products of YadA, a pYV-mediated fibrillar adhesin not encoded in frame by pCD. Carriage of pPCP by Y. pseudotuberculosis did not significantly influence virulence in mice. (+info)
The virulence plasmid-encoded impCAB operon enhances survival and induced mutagenesis in Shigella flexneri after exposure to UV radiation. (5/43669)Upon exposure to UV radiation, Shigella flexneri SA100 displayed survival and mutation frequencies comparable to those of Escherichia coli AB1157, which contains a functional UmuDC error-prone DNA repair system. Survival of SA100 after UV irradiation was associated with the presence of the 220-kb virulence plasmid, pVP. This plasmid encodes homologues of ImpA and ImpB, which comprise an error-prone DNA repair system encoded on plasmid TP110 that was initially identified in Salmonella typhimurium, and ImpC, encoded upstream of ImpA and ImpB. Although the impB gene was present in representatives of all four species of Shigella, not all isolates tested contained the gene. Shigella isolates that lacked impB were more sensitive to UV radiation than isolates that contained impB. The nucleotide sequence of a 2.4-kb DNA fragment containing the imp operon from S. flexneri SA100 pVP was 96% identical to the imp operon from the plasmid TP110. An SA100 derivative with a mutation in the impB gene had reduced survival following UV irradiation and less UV-induced mutagenesis relative to the parental strain. We also found that S. flexneri contained a chromosomally encoded umuDC operon; however, the umuDC promoter was not induced by exposure to UV radiation. This suggests that the imp operon but not the umuDC operon contributes to survival and induced mutagenesis in S. flexneri following exposure to UV radiation. (+info)
Molecular and evolutionary analysis of Borrelia burgdorferi 297 circular plasmid-encoded lipoproteins with OspE- and OspF-like leader peptides. (6/43669)We previously described two OspE and three OspF homologs in Borrelia burgdorferi 297 (D. R. Akins, S. F. Porcella, T. G. Popova, D. Shevchenko, S. I. Baker, M. Li, M. V. Norgard, and J. D. Radolf, Mol. Microbiol. 18:507-520, 1995; D. R. Akins, K. W. Bourell, M. J. Caimano, M. V. Norgard, and J. D. Radolf, J. Clin. Investig. 101:2240-2250, 1998). In this study, we characterized four additional lipoproteins with OspE/F-like leader peptides (Elps) and demonstrated that all are encoded on plasmids homologous to cp32 and cp18 from the B31 and N40 strains, respectively. Statistical analysis of sequence similarities using the binary comparison algorithm revealed that the nine lipoproteins from strain 297, as well as the OspE, OspF, and Erp proteins from the N40 and B31 strains, fall into three distinct families. Based upon the observation that these lipoproteins all contain highly conserved leader peptides, we now propose that the ancestors of each of the three families arose from gene fusion events which joined a common N terminus to unrelated proteins. Additionally, further sequence analysis of the strain 297 circular plasmids revealed that rearrangements appear to have played an important role in generating sequence diversity among the members of these three families and that recombinational events in the downstream flanking regions appear to have occurred independently of those within the lipoprotein-encoding genes. The association of hypervariable regions with genes which are differentially expressed and/or subject to immunological pressures suggests that the Lyme disease spirochete has exploited recombinatorial processes to foster its parasitic strategy and enhance its immunoevasiveness. (+info)
Reverse transcription-nested polymerase chain reaction for detecting p40 RNA of Borna disease virus, without risk of plasmid contamination. (7/43669)Several methods for the detection of Borna disease virus (BDV) RNA have been reported, one being the reverse transcription-nested polymerase chain reaction (RT-nested PCR) method. However, due to the possibility of contamination of the cloned DNA in a reaction tube, false-positive results might be obtained by RT-nested PCR. To detect only BDV RNA without anxiety of contamination, we developed an RT-nested PCR system using "mRNA selective PCR kit". Using this system, cDNA of BDV p40 in the plasmid (up to 5 x 10(7) molecules) was not amplified. BDV specific sequence was amplified from total RNA (more than 50 pg) of MDCK/BDV cells, which were persistently infected with BDV. These results indicate that this mRNA selective RT-nested PCR system can specifically amplify target RNA as distinguished from plasmid contaminated. (+info)
Diversity of rhizobia associated with Amorpha fruticosa isolated from Chinese soils and description of Mesorhizobium amorphae sp. nov. (8/43669)Fifty-five Chinese isolates from nodules of Amorpha fruticosa were characterized and compared with the type strains of the species and genera of bacteria which form nitrogen-fixing symbioses with leguminous host plants. A polyphasic approach, which included RFLP of PCR-amplified 16S rRNA genes, multilocus enzyme electrophoresis (MLEE), DNA-DNA hybridization, 16S rRNA gene sequencing, electrophoretic plasmid profiles, cross-nodulation and a phenotypic study, was used in the comparative analysis. The isolates originated from several different sites in China and they varied in their phenotypic and genetic characteristics. The majority of the isolates had moderate to slow growth rates, produced acid on YMA and harboured a 930 kb symbiotic plasmid (pSym). Five different RFLP patterns were identified among the 16S rRNA genes of all the isolates. Isolates grouped by PCR-RFLP of the 16S rRNA genes were also separated into groups by variation in MLEE profiles and by DNA-DNA hybridization. A representative isolate from each of these DNA homology groups had a separate position in a phylogenetic tree as determined from sequencing analysis of the 16S rRNA genes. A new species, Mesorhizobium amorphae, is proposed for the majority of the isolates, which belonged to a moderately slow- to slow-growing, acid-producing group based upon their distinct phylogenetic position, their unique electrophoretic type, their low DNA homology with reference strains representing the species within the genus Mesorhizobium and their distinct phenotypic features. Strain ACCC 19665 was chosen as the type strain for M. amorphae sp. nov. (+info)
The evolution of plasmid-carried antibiotic resistance | BMC Evolutionary Biology | Full Text
Antibiotic resistance represents a significant public health problem. When resistance genes are mobile, being carried on plasmids or phages, their spread can be greatly accelerated. Plasmids in particular have been implicated in the spread of antibiotic resistance genes. However, the selective pressures which favour plasmid-carried resistance genes have not been fully established. Here we address this issue with mathematical models of plasmid dynamics in response to different antibiotic treatment regimes. We show that transmission of plasmids is a key factor influencing plasmid-borne antibiotic resistance, but the dosage and interval between treatments is also important. Our results also hold when plasmids carrying the resistance gene are in competition with other plasmids that do not carry the resistance gene. By altering the interval between antibiotic treatments, and the dosage of antibiotic, we show that different treatment regimes can select for either plasmid-carried, or chromosome-carried,
Plasmids: Properties, Types and Functions - Learn Microbiology Online
Individual bacterial cells may contain several different types of plasmids and in some cases more than 10 at a time. Plasmids are generally isolated from the bacterial cells in the supercoiled configuration. So far, thousands of different types of plasmids have been isolated. More than 300 different types of naturally occurring plasmids have been isolated from E.coli alone. Though, plasmids are not considered as part of the cells genome, when a bacterial cell divides each daughter cells receives a copy of each plasmid. Plasmids can also be transferred from one bacterial cell to another by the process called conjugation. Plasmids that govern their own transfer by conjugation are called conjugative plasmids but not all plasmids are conjugative.. ...
Advantage of the F2A1B- IncF Pandemic Plasmid over IncC Plasmids in In Vitro Acquisition and Evolution of bla(CTX-M )Gene...
Despite a fitness cost imposed on bacterial hosts, large conjugative plasmids play a key role in the diffusion of resistance determinants, such as CTX-M extended-spectrum beta-lactamases. Among the large conjugative plasmids, IncF plasmids are the most predominant group, and an F2A1B- IncF-type plasmid encoding a CTX-M-15 variant was recently described as being strongly associated with the emerging worldwide Escherichia coli sequence type 131 (ST131)-O25bH4 H30Rx/C2 sublineage. In this context, we investigated the fitness cost of narrow-range F-type plasmids, including the F2A1B- IncF-type CTX-M-15 plasmid, and of broad-range C-type plasmids in the K-12-like J53-2 E. coli strain. Although all plasmids imposed a significant fitness cost to the bacterial host immediately after conjugation, we show, using an experimental-evolution approach, that a negative impact on the fitness of the host strain was maintained throughout 1,120 generations with the IncC-IncR plasmid, regardless of the presence or absence
Versatile low-copy-number plasmids for temperature-inducible overexpression of bacterial genes in Escherichia coli
Götting C, Thierbach G, Pühler A, Kalinowski J. Versatile low-copy-number plasmids for temperature-inducible overexpression of bacterial genes in Escherichia coli. BIOTECHNIQUES. 1998;24(3):362 ...
Major Families of Multiresistant Plasmids from Geographically and Epidemiologically Diverse Staphylococci | G3: Genes | Genomes...
The prevalence of 20-30 kb plasmids, almost half of which belong to only three restriction types by RFLP analysis, in staphylococci isolated from sources very distant in time and space suggests that these nonconjugative plasmids are surprisingly widespread for non-self-mobile plasmids. Plasmids in this size range can potentially be transferred by transducing phages (Lindsay and Holden 2006; Malachowa and Deleo 2010; Smillie et al. 2010); most phage genomes identified in staphylococci are ,40 kb, and transduction is thought to be restricted by phage genome size (Smillie et al. 2010). More of these 20-30 kb plasmids may be mobilizable than is apparent with current genome data if they contain mob genes not yet identified as such. However, the scarcity of conjugative plasmids (only 12 in total) implies that mobilization is rare and that staphylococcal plasmid transfer occurs mainly by transduction (Lindsay and Holden 2004; Lindsay 2010). The now larger dataset makes the mechanism of intercellular ...
Recombinant plasmid dna pfastbac-g2r-igg, which contains fragment of smallpox virus genome, coding factor of tumour necrosis,...
The presence of the amplified product PR is oraut by electrophoresis in 1% agarose gel.. Example 2. Construction of recombinant plasmid DNA pFastBac-G2R-IgG.. 5-10 μg of plasmid pFastBac  hydrolyzing the restriction endonucleases BamHI and HindIII; 1-5 μg of the amplified product corresponding gene G2R UPE without termination codon, hydrolyzing the restriction endonucleases BamHI and XbaI; the plasmid pBluescript-IgG1 hydrolyzing the restriction endonucleases XbaI and HindIII in standard conditions. The resulting fragments are electrophoresis in 1% agarose gel, followed by elution. 0.2 μg of the vector and 0.6 µg of fragments are ligated under standard conditions and received ligase mixture transform competent cells of E. coli strain XL-blue. Cellular Apr-clones grown at 37°C in LB medium containing 100 μg/ml ampicillin, until the stationary phase. Recombinant plasmid DNA secrete according to standard methods and analyzed by restriction endonucleases BamHI, XbaI and HindIII. Clones ...
Sequencing and comparative analysis of IncP-1 alpha antibiotic resistance plasmids reveal a highly conserved backbone and...
Although IncP-1 plasmids are important for horizontal gene transfer among bacteria, in particular antibiotic resistance spread, so far only three plasmids from the subgroup IncP-1 alpha have been completely sequenced. In this study we doubled this number. The three IncP-1 alpha plasmids pB5, pB11 and pSP21 were isolated from bacteria of two different sewage treatment plants and sequenced by a combination of next-generation and capillary sequencing technologies. A comparative analysis including the previously analysed IncP-1 alpha plasmids RK2, pTB11 and pBS228 revealed a highly conserved plasmid backbone (at least 99.9% DNA sequence identity) comprising 54 core genes. The accessory elements of the plasmid pB5 constitute a class 1 integron interrupting the parC gene and an IS6100 copy inserted into the integron. In addition, the tetracycline resistance genes tetAR and the ISTB11-like element are located between the klc operon and the trfA-ssb operon. Plasmid pB11 is loaded with a Tn5053-like ...
Investigation of diversity of plasmids carrying the blaTEM-52 gene - DTU Orbit
OBJECTIVES: To investigate the diversity of plasmids that carry blaTEM-52 genes among Escherichia coli and Salmonella enterica originating from animals, meat products and humans. METHODS: A collection of 22 blaTEM-52-encoding plasmids was characterized by restriction fragment length polymorphism (RFLP), replicon typing (by PCR or replicon sequencing), susceptibility testing, assessment of plasmid ability to self-transfer by conjugation and typing of the genetic environment of the blaTEM-52 gene. Detected IncI1 plasmids underwent further plasmid multilocus sequence typing. RESULTS: RFLP profiles demonstrated dissemination of blaTEM-52 in Denmark (imported meat from Germany), France, Belgium and the Netherlands from 2000 to 2006 by mainly two different plasmids, one encoding blaTEM-52b (IncX1A, 45 kb) and the other blaTEM-52c (IncI1, 80 kb). In addition, blaTEM-52b was also found to be located on various other plasmids belonging to IncA/C and IncL/M, while blaTEM-52c was found on IncN-like as well ...
Diverse Commensal Escherichia coli Clones and Plasmids Disseminate Antimicrobial Resistance Genes in Domestic Animals and...
In this semirural community, we found that numerically dominant commensal E. coli strains (showing similar antimicrobial resistance and same antibiotic resistance genes) colonizing children and domestic animals in the same period of time and in the same community are genotypically diverse. We also found that plasmids carrying the same antibiotic resistance genes were distinct, which is consistent with recent reports showing that AMR genes move frequently among different plasmids (28, 29). Our research suggests that a common pool of AMR genes could be cocirculating on different plasmids among different E. coli clones in a community (Table 2)-probably through dissemination mediated by transposons, integrons, or gene cassettes (28, 30). Even when the same resistance gene alleles and same plasmid replicon types were identified across isolates, the plasmids harboring these traits were still distinct. We also found potential evidence of Tn2 participation in mobility of the gene blaTEM-1B, as we found ...
Stress responses and replication of plasmids in bacterial cells | Microbial Cell Factories | Full Text
Plasmids, DNA (or rarely RNA) molecules which replicate in cells autonomously (independently of chromosomes) as non-essential genetic elements, play important roles for microbes grown under specific environmental conditions as well as in scientific laboratories and in biotechnology. For example, bacterial plasmids are excellent models in studies on regulation of DNA replication, and their derivatives are the most commonly used vectors in genetic engineering. Detailed mechanisms of replication initiation, which is the crucial process for efficient maintenance of plasmids in cells, have been elucidated for several plasmids. However, to understand plasmid biology, it is necessary to understand regulation of plasmid DNA replication in response to different environmental conditions in which host cells exist. Knowledge of such regulatory processes is also very important for those who use plasmids as expression vectors to produce large amounts of recombinant proteins. Variable conditions in large-scale
The IncP-1 plasmid backbone adapts to different host bacterial species and evolves through homologous recombination.
Plasmids are important members of the bacterial mobile gene pool, and are among the most important contributors to horizontal gene transfer between bacteria. They typically harbour a wide spectrum of host beneficial traits, such as antibiotic resistance, inserted into their backbones. Although these inserted elements have drawn considerable interest, evolutionary information about the plasmid backbones, which encode plasmid related traits, is sparse. Here we analyse 25 complete backbone genomes from the broad-host-range IncP-1 plasmid family. Phylogenetic analysis reveals seven clades, in which two plasmids that we isolated from a marine biofilm represent a novel clade. We also found that homologous recombination is a prominent feature of the plasmid backbone evolution. Analysis of genomic signatures indicates that the plasmids have adapted to different host bacterial species. Globally circulating IncP-1 plasmids hence contain mosaic structures of segments derived from several parental plasmids that
Plasmid partition system - Wikipedia
A plasmid partition system is a mechanism that assures the stable transmission of plasmids during bacterial cell division. Each plasmid has its independent replication system which controls the number of copies of the plasmid in a cell. The higher the copy number is, the more likely the two daughter cells will contain the plasmid. Generally, each molecule of plasmid diffuses randomly, so the probability of having a plasmid-less daughter cell is 21−N, where N is the number of copies. For instance, if there are 2 copies of a plasmid in a cell, there is a 50% chance of having one plasmid-less daughter cell. However, high-copy number plasmids have a cost for the hosting cell. This metabolic burden is lower for low-copy plasmids, but those have a higher probability of plasmid loss after a few generations. To control vertical transmission of plasmids, in addition to controlled-replication systems, bacterial plasmids use different maintenance strategies, such as multimer resolution systems, ...
Plasmid-mediated resistance - Wikipedia
Plasmid-mediated resistance is the transfer of antibiotic resistance genes which are carried on plasmids. The plasmids can be transferred between bacteria within the same species or between different species via conjugation. Plasmids often carry multiple antibiotic resistance genes, contributing to the spread of multidrug-resistance (MDR). Antibiotic resistance mediated by MDR plasmids severely limits the treatment options for the infections caused by Gram-negative bacteria, especially Enterobacteriaceae family. The global spread of MDR plasmids has been enhanced by selective pressure from antibiotic usage in human and veterinary medicine. Resistance plasmids by definition carry one or more antibiotic resistance genes. They are frequently accompanied by the genes encoding virulence determinants, specific enzymes or resistance to toxic heavy metals. Multiple resistance genes are commonly arranged in the resistance cassettes. The antibiotic resistance genes found on the plasmids confer resistance ...
What is the origin of plasmids? - Biology Stack Exchange
As I understand it, plasmids, like mitochondria, have their own genetic material and are capable of self-replication. According to Wikipedia: Plasmids are considered replicons, units of DNA capable of replicating autonomously within a suitable host. However, like viruses, they are not classified as life. Plasmids are transmitted from one bacterium to another through conjugation. Unlike viruses, plasmids are naked DNA. However, some classes of plasmids encode the conjugative sex pilus necessary for their own transfer. My understanding of that is that a bacteria gets their plasmids not because of the replication of their circular chromosome, nor because that chromosome have genes to code for the plasmid (I dont really know if thats possible), but because of the self-replication of their own plasmids. So, my question is how the first plasmid got into the first bacteria, if they are not in their chromosomes? Were they a virus other prokaryotic cell that had circular DNA, and got phagocytosed ...
NAVER Academic | Cloning ofCandida boidinii DNA fragments promoting autonomous replication of plasmids inSaccharomyces...
Fragments ofCandida boidinii chromosomal DNA were inserted into the integrative vector YIp-kanr and examined for the presence of sequences promoting autonomous replication of plasmids inSaccharomyces cerevisiae. Restriction maps of two plasmids, designated S6/4 and S6/5, originating from the sameS. cerevisiae transformant, were constructed. Southern hybridization data confirmed that the plasmids carry sequences from theC. boidinii chromosome. Both plasmids transformS. cerevisiae strains at 4-5-fold higher frequency than cloning vectors based on the replication origin of the 2μm plasmid. Mitotic stability of the constructed plasmids is similar to that of the 2μ-based vector pNF2 inS. cerevisiae.
A High Temperature-Sensitive Mutant of Synechococcus sp. PCC 7002 with Modifications in the Endogenous Plasmid, pAQ1 : Plant...
To study thermal adaptations in the cyanobacterium, Synechococcus sp. PCC 7002, we screened about 3,000 mutants for their tolerance to high temperature, and found one, SHT1, that is sensitive to high-temperature stress. The mutant had a modified gene construct in the endogenous plasmid, pAQ1. One of the four ORFs, ORF93, was duplicated, and its mRNA level was higher than in the wild type. At 38°C, the growth of SHT1 was retarded as compared with the wild type, and above 38°C, almost all the cells of SHT1 died. This temperature is much lower than that required for induction of heat shock proteins. Interestingly, in both the wild type and SHT1, the thermal stability of oxygen-evolving machinery increased upon acclimation to high temperatures. These findings indicate that the lack of thermal tolerance in the SHT1 strain is likely independent of the adaptation of the PSII complex and heat shock responses, whereas there are essential contributions of genes in the endogenous plasmid to the ...
Taxonomy and Epidemiology of Gram-negative Bacterial Plasmids Studied by DNA-DNA Filter Hybridization in Formamide |...
SUMMARY: Phylogenetic and epidemiological relatedness among transferable plasmids belonging to the IncC, IncM and IncH incompatibility groups has been studied by DNA-DNA filter hybridization. Hybridization was carried out on nitrocellulose microfilters, at low temperature, in formamide and under paraffin oil. The degree of hybridization among plasmids belonging to the IncC and IncM groups supported the conclusions drawn from genetic classification. Studies on relatedness among plasmids belonging to the IncH group allowed their classification into three phylogenetic sub-groups. Comparison of DNA sequences of three plasmids sharing the same genetic properties and isolated from different bacterial species suggested an epidemiological spread of the same plasmid.
Recreate original plasmid by cutting out insert - Molecular Cloning - BioForum
Recreate original plasmid by cutting out insert - posted in Molecular Cloning: Maybe this is a stupid question, but I am going to ask it anyway: I would like to cut out an insert from my plasmid (pET28c) between restriction sites Nhe1 and Not1 and ligate the plasmid back to its orginial state without the insert (no I dont have the original plasmid anymore). However, obviously these restriction sites dont generate compatible sticky ends. Does anyone know how to recreate this...
Complete genetic analysis of plasmids carrying mcr-1 and other resistance genes in an Escherichia coli isolate of animal origin...
Objectives: To investigate the genetic features of three plasmids recovered from an MCR-1 and ESBL-producing Escherichia coli strain, HYEC7, and characterize the transmission mechanism of mcr-1. Methods: The genetic profiles of three plasmids were determined by PCR, S1-PFGE, Southern hybridization and WGS analysis. The ability of the mcr-1-bearing plasmid to undergo conjugation was also assessed. The mcr-1-bearing transposon Tn6330 was characterized by PCR and DNA sequencing. Results: Complete sequences of three plasmids were obtained. A non-conjugative phage P7-like plasmid, pHYEC7-mcr1, was found to harbour the mcr-1-bearing transposon Tn6330, which could be excised from the plasmid by generating a circular intermediate harbouring mcr-1 and the ISApl1 element. The insertion of the circular intermediate into another plasmid, pHYEC7-IncHI2, could form pHNSHP45-2, the original IncHI2-type mcr-1-carrying plasmid that was reported. The third plasmid, pHYEC7-110, harboured two replicons, IncX1 and ...
Natural selection, infectious transfer, and the existence conditions for bacterial plasmids
Despite the near-ubiquity of plasmids in bacterial populations and the profound contribution of plasmid-borne genes and infectious gene transfer to the adaptation and evolution of bacteria, the mechanisms responsible for the maintenance of plasmids in bacterial populations are poorly understood. In this report, we address the question of how plasmids manage to persist over evolutionary time. Previous explanations have typically relied upon the ability of plasmids to deliver occasionally-useful genes to the right place at the right time. In contrast, we present a general mathematical proof that if (as suggested by several empirical studies) plasmids are not infectiously transmitted at a rate high enough to be maintained as genetic parasites in single populations, they will not be able to persist indefinately in these populations by carrying genes that are beneficial or sometimes beneficial to their host bacteria. Using more specific mathematical models, along with computer simulations, we ...
uncut plasmid dna vs linearized plasmid gel - Molecular Cloning - BioForum
uncut plasmid dna vs linearized plasmid gel - posted in Molecular Cloning: Hello, I am going to run a gel comparing my uncut plasmid dna vs linearized plasmid. My insert is 135 bps and my vector is 3Kb. What can I expect to see on my gel, and how many bands can I expect to see. I am assuming the uncut plasmid will have several bands at different sizes and the linearized will have only one, is that right ? I am assuming the total size of my product will now be 3135 bps. Pls advise.
3.6 Plasmid Cloning Vectors for E. coli
Figure 20. The pBR322 plasmid. Two genes of pBR322 confer resistance to antibiotics to any cell that contains the plasmid. AmpR confers resistance to amplicillin and tetR confers resistance to tetracycline to cells containing the plasmids. AmpR is a gene that codes for the periplasmic enzyme beta-lactamase that cleaves the ring structure found in amphicillin, which is a penicillin antibiotic. TetR is a gene that codes for a protein that modifies the bacterial cell wall and prevents tetracycline from entering the cell.. Multiple restriction endonuclease sites are present where foreign DNA fragments may be inserted.. Relaxed plasmid DNA replication continues in the presence of chloramphenicol. An interesting feature of this plasmid is that relaxed plasmid DNA replication continues even in the presence of an inhibitor of protein synthesis such as chloramphenicol. This feature allows increased yields of plasmid/cell of up to 100-fold ...
Yeast Plasmid Miniprep - Yeast Plasmid Isolation Kit
Easy Yeast Plasmid Isolation Kit: rescue plasmids from yeast using spin columns. Simple and highly efficient method. Zymolyase included.
Yeast Plasmid Miniprep - Yeast Plasmid Isolation Kit
Easy Yeast Plasmid Isolation Kit: rescue plasmids from yeast using spin columns. Simple and highly efficient method. Zymolyase included.
ASMscience | Resolution of Multimeric Forms of Circular Plasmids and Chromosomes
One of the disadvantages of circular plasmids and chromosomes is their high sensitivity to rearrangements caused by homologous recombination. Odd numbers of crossing-over occurring during or after replication of a circular replicon result in the formation of a dimeric molecule in which the two copies of the replicon are fused. If they are not converted back to monomers, the dimers of replicons may fail to correctly segregate at the time of cell division. Resolution of multimeric forms of circular plasmids and chromosomes is mediated by site-specific recombination, and the enzymes that catalyze this type of reaction fall into two families of proteins: the serine and tyrosine recombinase families. Here we give an overview of the variety of site-specific resolution systems found on circular plasmids and chromosomes.
Santa Cruz Biotechnology now offers target-specific CRISPR/Cas9 Knockout (KO) Plasmids, CRISPR Double Nickase Plasmids and CRISPR/dCas9 Activation Plasmids for over 18,910 human and 18,340 mouse protein encoding genes. CRISPR/Cas9 KO Plasmids and CRISPR Double Nickase Plasmids enable the identification and cleavage of specific genes encoding a protein of interest thereby eliminating production of that gene product (protein). CRISPR/dCas9 Activation Plasmids activate endogenous gene transcription using a robust synergistic activation mediator (SAM) system. This exciting new technology is a useful tool for studying protein function and signalling pathways ...
Monitoring the Dissemination of the Broad-Host-Range Plasmid pB10 in Sediment Microcosms by Quantitative PCR - Université de...
Studying the transfer of specific mobile genetic elements in complex environmental matrices remains difficult because suitable molecular tools are not yet available to back up classical culture-dependent approaches. In this report, we show that quantitative PCR could be used to monitor the dissemination of the broad-host-range plasmid pB10 in sediment microcosms. This approach lies in the differential measurement of the host and plasmid DNAs used to inoculate the microcosms, using a particular design of quantitative PCR primers/probes where we took advantage of the mosaic aspect of the bacterial genomes to achieve a highly specific quantitative PCR detection system.
The partition system of multidrug resistance plasmid TP228 includes a novel protein that epitomizes an evolutionarily distinct...
The segregational stability of bacterial, low-copy-number plasmids is promoted primarily by active partition. The plasmid-specified components of the prototypical P1 plasmid partition system consist of two proteins, ParA (44.3 kDa) and ParB (38.5 kDa), which, in conjunction with integration host fac …
Spatial structure and nutrients promote invasion of IncP-1 plasmids in bacterial populations
In spite of the importance of plasmids in bacterial adaptation we have a poor understanding of their dynamics. It is not known if or how plasmids persist in and spread through (invade) a bacterial population when there is no selection for plasmid-encoded traits. Moreover, the differences in dynamics between spatially structured and mixed populations are poorly understood. Through a joint experimental/theoretical approach we tested the hypothesis that self-transmissible IncP-1 plasmids can invade a bacterial population in the absence of selection when initially very rare, but only in spatially structured habitats and when nutrients are regularly replenished. Using protocols that differed in degree of spatial structure and nutrient levels, the invasiveness of plasmid pB10 in E. coli was monitored during at least 15 days, with an initial fraction of plasmid-bearing (p+) cells as low as 1E-7. To further explore the mechanisms underlying plasmid dynamics we developed a spatially explicit mathematical ...
Manganese in the structure of Crystal Structure of the Replication Initiator Protein Encoded on Plasmid PMV158 (Repb), Trigonal...
Mono- and Stereopictres of 5.0 Angstrom coordination sphere of Manganese atom in PDB 3dkx: Crystal Structure of the Replication Initiator Protein Encoded on Plasmid PMV158 (Repb), Trigonal Form, to 2.7 Ang Resolution
Novel Plasmid Technology at PEGS Boston | Batavia Biosciences
Difficult to Express Proteins: Novel Plasmid Technology to Significantly Increase Product Yield in CHO Cells, by Marco Cacciutolo
Key features of mcr-1 -bearing plasmids from Escherichia coli isolated from humans and food | Antimicrobial Resistance &...
Mcr-1-harboring Enterobacteriaceae are reported worldwide since their first discovery in 2015. However, a limited number of studies are available that compared full-length plasmid sequences of human and animal origins. In this study, mcr-1-bearing plasmids from seven Escherichia coli isolates recovered from patients (n = 3), poultry meat (n = 2) and turkey meat (n = 2) in Switzerland were further analyzed and compared. Isolates were characterized by multilocus sequence typing (MLST). The mcr-1-bearing plasmids were transferred by transformation into reference strain E. coli DH5α and MCR-1-producing transformants were selected on LB-agar supplemented with 2 mg/L colistin. Purified plasmids were then sequenced and compared. MLST revealed six distinct STs, illustrating the high clonal diversity among mcr-1-positive E. coli isolates of different origins. Two different mcr-1-positive plasmids were identified from a single E. coli ST48 human isolate. All other isolates possessed a
Other Plasmid Components
The reason that we stock plasmids that only contain one terminator is because sometimes, despite the increased versatility, researchers may not want unnecessary terminators in their plasmids, for example when size constraints are an issue. The button below takes you to the plasmids with single terminators. If you prefer to use a plasmid with triple terminator, we suggest you search using the Plasmid Seach tool since nearly all our plasmids have triple terminators. The Plasmid Search tool will give you access to our full plasmid catalogue.. Finally, please try the Design Your Own Plasmid Online button below, to see what our cloning system can do for you. And remember that, while our plasmids are designed for easy cloning modifications, we are happy to do it for you if you prefer to outsource the cloning work.. ...
High-resolution mapping of plasmid transcriptomes in different host bacteria | BMC Genomics | Full Text
Prokaryotic transcriptomes change not only in response to physiological parameters but also to genetic rearrangements mediated by mobile elements. Plasmids are extrachromosomal genetic elements that replicate autonomously, and many can be transmitted between different strains through conjugation. Plasmids provide benefits to their hosts, such as resistance to antibiotics or degradation of recalcitrant aromatic compounds ; however, in several cases, the carriage of a large plasmid results in changes in the transcriptome of the host chromosome [2-4]. Similar to the effects of plasmid carriage on the transcriptional network of the host chromosome, differences in host background can alter the transcription patterns of backbone and accessory genes on a plasmid. Many plasmid backbone genes essential for conjugative transfer, replication initiation, and active partitioning are regulated both autogenously and by host factors . Additionally, a number of plasmid-encoded degradative accessory genes ...
CRIMoClo plasmids for modular assembly and orthogonal chromosomal integration of synthetic circuits in Escherichia coli |...
Synthetic biology heavily depends on rapid and simple techniques for DNA engineering, such as Ligase Cycling Reaction (LCR), Gibson assembly and Golden Gate assembly, all of which allow for fast, multi-fragment DNA assembly. A major enhancement of Golden Gate assembly is represented by the Modular Cloning (MoClo) system that allows for simple library propagation and combinatorial construction of genetic circuits from reusable parts. Yet, one limitation of the MoClo system is that all circuits are assembled in low- and medium copy plasmids, while a rapid route to chromosomal integration is lacking. To overcome this bottleneck, here we took advantage of the conditional-replication, integration, and modular (CRIM) plasmids, which can be integrated in single copies into the chromosome of Escherichia coli and related bacteria by site-specific recombination at different phage attachment (att) sites. By combining the modularity of the MoClo system with the CRIM plasmids features we created a set of 32 novel
ID YEP367 preliminary; circular DNA; SYN; 8400 BP. XX AC ATCC37735; XX DT 01-JUL-1993 (Rel. 7, Created) DT 01-JUL-1995 (Rel. 12, Last updated, Version 1) XX DE Saccharomyces/E.coli plasmid vector YEp367 - incomplete. XX KW cloning vector. XX OS Cloning vector OC Artificial sequences; Cloning vehicles. XX RN  RC YEp352E from YEp352 & linker RC YEp363A from pNM480 & YEp351 RC YEp353A from pNM480 & YEp352 RC YEp353 from YEp353A & YEp352E RC YEp354A from pNM481 & YEp352 RC YEp354 from YEp354A & YEp352E RC YEp355A from pNM482 & YEp352 RC YEp355 from YEp355A & YEp352E RC YEp356, YEp356R from YEp353 & pUC18 RC YEp357, YEp357R from YEp354 & pUC18 RC YEp358, YEp358R from YEp355 & pUC18 RC YEp363 from YEp363A & YEp353 RC YEp364 from YEp363A & YEp354 RC YEp365 from YEp363A & YEp355 RC YEp366 from YEp363A & YEp356 RC YEp367 from YEp363A & YEp357 RC YEp368 from YEp363A & YEp358 RC YEp366R from YEp363A & YEp356R RC YEp367R from YEp363A & YEp357R RC YEp368R from YEp363A & YEp358R RC YIp353 from YEp353 & ...
Background Prokaryotic plasmids have played out significant roles in the evolution - Suppression of KRas-mutant cancer through...
Background Prokaryotic plasmids have played out significant roles in the evolution of bacterial genomes and have a great impact on the metabolic functions of the host cell. variable genes (distributed genes and unique genes) than to the chromosomal core genes. Although all the functional categories of the chromosomal genes were exhibited by the plasmid genes, the proportions of each category differed between these two gene sets. The 598 gene families shared between chromosomes and plasmids displayed a uniform distribution between the two groups. A phylogenetic analysis of the shared genes, including the chromosomal core gene set, indicated that gene exchange events between plasmids and chromosomes occurred frequently during the evolutionary histories of the strains and species in this group. Moreover, the shared genes between plasmids and chromosomes usually had different promoter and terminator sequences, suggesting that they are regulated by different elements at the transcriptional level. ...
Plasmids in Bacteria
www.MOLUNA.de Plasmids in Bacteria  - Structure and Evolution.- Plasmids as Organisms.- Report on a Workshop: Structure and Function.- Evolutionary Relevance of Genetic Rearrangements Involving Plasmids.- Mechanisms of Transposition in Bacteria.- Insertion of Transcriptional Elements Outside the Replication Region Can Interfere with Replication, Maintenance, and Stability of ColE1-Derived Plasmids.- Studies on the Transposition of IS1.- On
ID YEP353 preliminary; circular DNA; SYN; 7944 BP. XX AC U03500; ATCC37725; XX DT 01-JUL-1993 (Rel. 7, Created) DT 01-JUL-1995 (Rel. 12, Last updated, Version 1) XX DE Saccharomyces/E.coli plasmid vector YEp353 - complete. XX KW cloning vector. XX OS Cloning vector OC Artificial sequences; Cloning vehicles. XX RN  RP 1-7944 RC YEp352E from YEp352 & linker RC YEp363A from pNM480 & YEp351 RC YEp353A from pNM480 & YEp352 RC YEp353 from YEp353A & YEp352E RC YEp354A from pNM481 & YEp352 RC YEp354 from YEp354A & YEp352E RC YEp355A from pNM482 & YEp352 RC YEp355 from YEp355A & YEp352E RC YEp356, YEp356R from YEp353 & pUC18 RC YEp357, YEp357R from YEp354 & pUC18 RC YEp358, YEp358R from YEp355 & pUC18 RC YEp363 from YEp363A & YEp353 RC YEp364 from YEp363A & YEp354 RC YEp365 from YEp363A & YEp355 RC YEp366 from YEp363A & YEp356 RC YEp367 from YEp363A & YEp357 RC YEp368 from YEp363A & YEp358 RC YEp366R from YEp363A & YEp356R RC YEp367R from YEp363A & YEp357R RC YEp368R from YEp363A & YEp358R RC YIp353 ...
DNA plasmids, AFM - Stock Image G110/0752 - Science Photo Library
DNA plasmids. Coloured atomic force micrograph of numerous pGL3 plasmids of DNA (deoxyribonucleic acid). A plasmid is a loop of DNA that can exist in a cell independently of the cells chromosomal DNA. Plasmids are important as they can replicate and express genes despite being outside the organisms chromosomes. This means that they can be used to introduce genes to organisms, a process used in gene therapy and genetic engineering. Genes, specific lengths of DNA, determine the development and characteristics of an organism. Atomic force microscopes make an image by moving a sensitive probe over a surface. Magnification: x24,000 at 6x7cm size. - Stock Image G110/0752
Transitory plasmid instability during the exponential phase of growth of a recombinant Escherichia coli | SpringerLink
The present results concern the recombinant bacteria Escherichia coliHB101(GAPDH) which produces glyceraldehyde 3-phosphate dehydrogenase. An unusual phenomenon was noticed concerning the plasmid...
Evolutionary dynamics on multicopy plasmids | Seminars
Many bacteria carry extra DNA molecules beyond their chromosome, so-called plasmids. While plasmids are apriori costly to the cell, they ...
Berkeley2006-PlasmidPartsMain - 2006.igem.org
The combination of a p15A and a colE1 origin of replication plasmid is the most common two-plasmid system for use in E. coli. The pSB1A* series of plasmids contain the colE1 plasmid and cannot be co-maintained at stable copy numbers in combination due to plasmid incompatibility. Here, two plasmids in the same cell with origins from the same incompatibility group compete for replication machinery, resulting in unpredictable copy number and often the exclusion of one of the plasmids. This is circumvented by cotransforming plasmids with origins from distinct incompatibility groups. Since no p15A origin plasmid was present in the Registry, we developed part J23001, or pSB3C6 for our studies. This plasmid was used for all our riboregulator experiments for the locked-RFP reporter. Shown here is the variant of the plasmid containing part J01022. Plasmids for constructing basic Biobrick parts ...
Comparative analysis of conjugative plasmids mediating gentamicin resistance in Staphylococcus aureus<...
TY - JOUR. T1 - Comparative analysis of conjugative plasmids mediating gentamicin resistance in Staphylococcus aureus. AU - Goering, R. V.. AU - Ruff, E. A.. PY - 1983/1/1. Y1 - 1983/1/1. N2 - Five gentamicin-resistant clinical isolates of Staphylococcus aureus were found to contain self-transmissible plasmids of 32 to 37 megadaltons in size. Restriction endonuclease digests of the plasmids were markedly similar to those of reference plasmids of unrelated geographical origin, thus suggesting the significant contribution of common conjugal plasmids to the emergence of gentamicin resistance in S. aureus populations.. AB - Five gentamicin-resistant clinical isolates of Staphylococcus aureus were found to contain self-transmissible plasmids of 32 to 37 megadaltons in size. Restriction endonuclease digests of the plasmids were markedly similar to those of reference plasmids of unrelated geographical origin, thus suggesting the significant contribution of common conjugal plasmids to the emergence of ...
Recombination of plasmids in a carbapenem-resistant NDM-5-producing clinical Escherichia coli isolate - CityU Scholars | A...
Objectives: To investigate the genetic features of five plasmids recovered from an NDM-5-producing clinical Escherichia coli strain, BJ114, and to characterize the plasmid recombination event that occurred during the conjugation process. Methods: The genetic profiles of the five plasmids were determined by PCR, conjugation, S1-PFGE, Southern hybridization and WGS analysis. Plasmid sequences were analysed with various bioinformatic tools. Results: Complete sequences of five plasmids were obtained. Two small plasmids, pBJ114-141 and pBJ114-46, were speculated to have recombined into a large fusion plasmid, pBJ114T-190. When conjugated to other E. coli strains, some of the fusion plasmids were able to be resolved into the original two single plasmids. A nonconjugative plasmid, pBJ114-96, exhibited a high degree of sequence identity with the phage P7-like plasmid as well as an mcr-1-bearing plasmid. Another plasmid, pBJ114-78, was found to contain multidrug resistance genes and various mobile ...
Going around in circles: Virulence plasmids in enteric pathogens - Immunology
© 2018 Macmillan Publishers Ltd., part of Springer Nature. Plasmids have a major role in the development of disease caused by enteric bacterial pathogens. Virulence plasmids are usually large (|40 kb) low copy elements and encode genes that promote host-pathogen interactions. Although virulence plasmids provide advantages to bacteria in specific conditions, they often impose fitness costs on their host. In this Review, we discuss virulence plasmids in Enterobacteriaceae that are important causes of diarrhoea in humans, Shigella spp., Salmonella spp., Yersinia spp and pathovars of Escherichia coli. We contrast these plasmids with those that are routinely used in the laboratory and outline the mechanisms by which virulence plasmids are maintained in bacterial populations. We highlight examples of virulence plasmids that encode multiple mechanisms for their maintenance (for example, toxin-antitoxin and partitioning systems) and speculate on how these might contribute to their propagation and success.
Plasmid addiction systems : Perspectives and applications in biotechnology
Biotechnical production processes often operate with plasmid-based expression systems in well-established prokaryotic and eukaryotic hosts such as Escherichia coli or Saccharomyces cerevisiae, respectively. Genetically engineered organisms produce important chemicals, biopolymers, biofuels and high-value proteins like insulin. In those bioprocesses plasmids in recombinant hosts have an essential impact on productivity. Plasmid-free cells lead to losses in the entire product recovery and decrease the profitability of the whole process. Use of antibiotics in industrial fermentations is not an applicable option to maintain plasmid stability. Especially in pharmaceutical or GMP-based fermentation processes, deployed antibiotics must be inactivated and removed. Several plasmid addiction systems (PAS) were described in the literature. However, not every system has reached a full applicable state. This review compares most known addiction systems and is focusing on biotechnical applications.. ...
Restriction endonuclease analysis of Staphylococcus aureus plasmid DNA from three continents<...
TY - JOUR. T1 - Restriction endonuclease analysis of Staphylococcus aureus plasmid DNA from three continents. AU - Doebbeling, Bradley. AU - Pfaller, M. A.. AU - Hollis, R. J.. AU - Boyken, L. D.. AU - Pignatari, A. C.. AU - Herwaldt, L. A.. AU - Wenzel, R. P.. PY - 1992/1. Y1 - 1992/1. N2 - Staphylococcus aureus isolates (n=1201) from 20 centers in Europe, the USA and Brazil were evaluated for the presence of epidemiologic markers. Plasmid typing and restriction endonuclease analysis of plasmid DNA confirmed the presence of an apparently identical plasmid in 13 % of clinical isolates. The plasmid was recovered from all 20 hospitals studied, with an overall frequency of ,10 % on each of the three continents. Since relatively few staphylococcal plasmids may be shared by epidemiologically unrelated strains, there are inherent limitations to this otherwise useful technique. Additionally, these data demonstrate the importance of including unrelated strains of Staphylococcus aureus from the local ...
Frontiers | Identification of Novel Conjugative Plasmids with Multiple Copies of fosB that Confer High-Level Fosfomycin...
To further characterize the fosB-carrying plasmids of 19 vancomycin-resistant enterococci, the complete sequences of the fosB- and vanA-containing plasmids of E. faecium (pEMA120) and E. avium (pEA19081) were obtained by single-molecule, real-time sequencing. We found that these two plasmids are essentially identical (99.99% nucleotide sequence identity), which proved the possibility of interspecies transmission. Comparative analysis of the plasmids revealed that the backbone of pEMA120 is 99% similar to a conjugative fosB-negative E. faecium plasmid, pZB18. There is a traE disrupted in the transfer region of pEMA120, in comparison to pZB18 with an intact traE. The difference of their transfer frequencies between pEMA120 and pZB18 suggests this interruption of traE might affect conjugative transfer. Two copies of the fosB gene linked to a tnpA gene, forming an ISL3-like transposon, were found at separate locations within pEMA120, which had not been reported previously. These two fosB-carrying
Molecular analysis of the replication region of the pCIZ2 plasmid from the multiple bacteriocin producer strain Enterococcus...
The sequence analysis of the 7383 bp plasmid pCIZ2 from |i|Enterococcus faecium|/i| L50 enabled the identification of a DNA region involved in its replication. The structural organization of the pCIZ2 replication region is highly similar to those of well-known theta-replicating plasmids. It contains an untranslated region, the putative replication origin (|i|ori|/i|), constituted by two sets of direct repeats of 12 and 22 bp (iterons), and followed by three open-reading frames (|i|orf8|/i| to |i|orf10|/i|). |i|orf8|/i| encodes the replication initiation protein (RepE). The transcriptional start site of the replication locus was identified 13 nucleotides upstream of the |i|repE|/i| start codon. A two-dimensional agarose gel electrophoresis analysis revealed pCIZ2 intermediates profile typical of the theta-type replication mechanism. Subcloning of different DNA fragments of the pCIZ2 replication region in |i|Escherichia coli|/i| and, subsequently, in the plasmidless |i|E. faecium|/i| L50/14-2 allowed the
Vývoj plazmidů - interakce plazmidů rezistence a virulence u Salmonella enterica - Mgr. Tereza Kubasová, Ph.D.
Gene transfer mediated by plasmids plays an important role in the spread of antibiotic resistance genes. Nowadays, high molecular weight plasmids carrying resistance genes to antibiotics and growth inhibitors are commonly found in pathogenic bacteria. Moreover, conjugative plasmids coding for antibiotic resistance may interact with virulence plasmids thus resulting in plasmid transferring both antibiotic resistance and virulence genes. In this thesis I therefore pursued genomics and proteomics studies on plasmids of two incompatibility group. In the first part of the thesis we determined the complete nucleotide sequences of multidrug resistance plasmid p9134 and its two variants, p9134dT which spontaneously lost tetracycline resistance gene, and p9134dAT which spontaneously lost ampicillin and tetracycline resistance genes. Plasmids were 133 802 bp, 127 291 bp a 109 512 bp in size, respectively, and their basic backbone was similar to that of IncI plasmids. In plasmid p9134 genes coding for ...
Evolution of IncA/C blaCMY-2-Carrying Plasmids by Acquisition of the blaNDM-1 Carbapenemase Gene | Antimicrobial Agents and...
ii) Evolution of the blaNDM-1 locus.The mer operon of pAR060302 is part of a complex hybrid transposon inserted into the IncA/C backbone flanked by direct repeats, and related structures are found in other IncA/C plasmids. The strA-strB genes in pYR1 are part of a Tn5393 that was recently described (3, 4). The synteny among the compared plasmids restarted at the DNA primase gene (Fig. 1). IncA/C plasmid pP91278 from P. damselae likely represents the IncA/C scaffold before the acquisition of the accessory genes, since it does not show any insertion between the phage-integrase, rhs, and DNA primase genes (Fig. 1).. An analysis of the pNDM-KN sequence revealed that the rhs gene was deleted at its 3′ and 5′ extremities and was adjacent to the heat shock chaperone groEL-groES cluster (28). In pAR060302, this cluster flanked a class 1 integron that contained the aac(3)-IVa and aadA1 gene cassettes, but it was absent in plasmids pYR1 and pP91278 (Fig. 1). Analysis of the sequences located at the ...
[email protected]: The Bacillus subtilis conjugative transposon ICEBs1 mobilizes plasmids lacking dedicated...
Integrative and conjugative elements (ICEs, also known as conjugative transposons) are mobile elements that are found integrated in a host genome and can excise and transfer to recipient cells via conjugation. ICEs and conjugative plasmids are found in many bacteria and are important agents of horizontal gene transfer and microbial evolution. Conjugative elements are capable of self-transfer and also capable of mobilizing other DNA elements that are not able to self-transfer. Plasmids that can be mobilized by conjugative elements are generally thought to contain an origin of transfer (oriT), from which mobilization initiates, and to encode a mobilization protein (Mob, a relaxase) that nicks a site in oriT and covalently attaches to the DNA to be transferred. Plasmids that do not have both an oriT and a cognate mob are thought to be nonmobilizable. We found that Bacillus subtilis carrying the integrative and conjugative element ICEBs1 can transfer three different plasmids to recipient bacteria at ...
Complete Sequencing of Plasmids ContainingblaOXA-163andblaOXA-48in Escherichia coli Sequence Type 131 - Wellcome Centre for...
|jats:title|ABSTRACT|/jats:title||jats:p|OXA-48-like enzymes have emerged as important extended-spectrum β-lactamases/carbapenemases in|jats:named-content xmlns:xlink=http://www.w3.org/1999/xlink content-type=genus-species xlink:type=simple|Escherichia coli|/jats:named-content|sequence type 131 (ST131). We report the structures of the first fully sequenced|jats:italic|bla|/jats:italic||jats:sub|OXA-163|/jats:sub|plasmid and of two other|jats:italic|bla|/jats:italic||jats:sub|OXA-48|/jats:sub|plasmids in this lineage.|jats:italic|bla|/jats:italic||jats:sub|OXA-163|/jats:sub|was located on a 71-kb IncN plasmid with other resistance genes.|jats:italic|bla|/jats:italic||jats:sub|OXA-48|/jats:sub|was present on IncL/M plasmids, genetically similar to other|jats:italic|bla|/jats:italic||jats:sub|OXA-48|/jats:sub|plasmid sequences, and consistent with interspecies/interlineage spread. The presence of|jats:italic|bla|/jats:italic||jats:sub|OXA-48-like|/jats:sub|genes on epidemic plasmids in ST131 is of
ASMscience | Plasmids as Genetic Tool
This chapter provides a broad overview of many applications of plasmids for genetic analysis, primarily in bacteria. Ever since DNA sequencing became accessible to most research laboratories, reverse genetic analysis has become a standard experimental approach to study bacterial gene function. Similar suicide vectors have also been used for nontargeted insertional mutagenesis by cloning random chromosomal DNA fragments into the plasmid. The use of suicide vectors also allows for easy identification of the insertion mutations. Plasmids that utilize different combinations of double-counter selective markers have been used for diverse applications, including the search for extremely rare suppressor mutations of essential Escherichia coli genes, and to improve the efficiency of allelic exchange on bacterial artificial chromosomes (BACs). Although temperature-sensitive vectors represent the majority of conditionally replicating plasmids, other plasmids that exhibit conditional replication have been described
Detection of plasmid DNA vectors following gene transfer to the murine airways. - Radcliffe Department of Medicine
Non-viral gene therapy is being considered as a treatment for cystic fibrosis. In clinical studies and in studies using the mouse airways as a model, current formulations result in only transient transgene expression. A number of reasons for this have been proposed including the loss of plasmid DNA from cells. The aim of these studies was to investigate why transgene expression from non-viral vectors is transient in the mouse lung. Plasmid DNA encoding the luciferase reporter gene was complexed with the cationic lipid GL67 and delivered to the mouse airways. The persistence of plasmid DNA in the mouse lungs was investigated using quantitative PCR and Southern hybridization. Results showed that intact plasmid DNA persisted in the mouse lung in the absence of any detectable luciferase activity. The de novo methylation of plasmid DNA in vivo was investigated as a potential cause of this transient gene expression but results suggested that plasmid DNA does not become de novo methylated in the mouse lung.
Frequency-dependent advantages of plasmid carriage by Pseudomonas in homogeneous and spatially structured environments. -...
The conditions promoting the persistence of a plasmid carrying a trait that may be mutually beneficial to other cells in its vicinity were studied in structured and unstructured environments. A large plasmid encoding mercury resistance in Pseudomonas fluorescens was used, and the mercury concentration allowing invasion from rare for both plasmid-bearing and plasmid-free cells was determined for different initial inoculum densities in batch-culture structured (filter surface) and unstructured (mixed broth) environments. A range of mercury concentrations were found where both cell types could coexist, the regions being relatively similar in the two types of environment although density-dependent in the unstructured environment. The coexistence is explained in terms of frequency-dependent selection of the mutually beneficial mercury resistance trait, and the dynamics of bacterial growth under batch culture conditions. However, the region of coexistence was complicated by conjugation which increased plasmid
International Society for Plasmid Biology | International Society for Plasmids and other Mobile Genetic Elements
The International Society for Plasmid Biology and other Mobile Genetic Elements (ISPB) was established in 2004. The purpose of the ISPB is to promote the study of plasmids and other mobile genetic elements.. The Society will engage in collective activities that increase the awareness and understanding of the importance of these genetic elements both in the scientific community and in the public arena.. The Society will highlight important discoveries in the plasmid field as well as new and novel applications of plasmids and other mobile genetic elements, and will act as an advocate for increased research support of this field.. ISPB will also promote the importance of retaining the study of plasmids and other mobile genetic elements in undergraduate teaching curriculum.. The Society will provide a forum for the plasmid biology community to work with sequence databases to ensure the highest quality of annotation.. The constitution of ISPB can be found here.. Information about previous conferences ...
The Isolation of Strains of Bacillus subtilis Showing Improved Plasmid Stability Characteristics by Means of Selective...
SUMMARY: A PUB 110-derived plasmid encoding chloramphenicol resistance, kanamycin resistance and high-temperature α-amylase showed a high degree of segregational instability when inserted into Bacillus subtilis. In an attempt to obtain stable derivatives, the organism was grown in chemostat culture in the presence of chloramphenicol. It was periodically found necessary to increase the concentration of chloramphenicol in the medium feed in order to avoid plasmid loss. Strains were isolated after 19 and 160 generations, which showed high levels of plasmid stability. This characteristic appeared to be genotypic. No detectable difference in plasmid copy number was found between the original and the improved strains. The stability characteristics resided in the host, rather than in the plasmid. Stable isolates possessed elevated MICs for both chloramphenicol and kanamycin. Their maximum specific growth rates were higher than that of the original strain, and similar to that of the plasmid-free parent strain.
Comparison of Electrically Mediated and Liposome-Complexed Plasmid DNA by Loree C. Heller, Mark J. Jaroszeski et al.
BACKGROUND: Electroporation is an established technique for enhancing plasmid delivery to many tissues in vivo, including the skin. We have previously demonstrated efficient delivery of plasmid DNA to the skin utilizing a custom-built four-plate electrode. The experiments described here further evaluate cutaneous plasmid delivery using in vivo electroporation. Plasmid expression levels are compared to those after liposome mediated delivery. METHODS: Enhanced electrically-mediated delivery, and less extensively, liposome complexed delivery, of a plasmid encoding the reporter luciferase was tested in rodent skin. Expression kinetics and tissue damage were explored as well as testing in a second rodent model. RESULTS: Experiments confirm that electroporation alone is more effective in enhancing reporter gene expression than plasmid injection alone, plasmid conjugation with liposomes followed by injection, or than the combination of liposomes and electroporation. However, with two time courses of multiple
Transmission of IncN Plasmids Carrying blaCTX-M-1 between Commensal Escherichia coli in Pigs and Farm Workers | Antimicrobial...
Extended-spectrum cephalosporins have been classified by the World Health Organization as critically important antibiotics in human medicine (19). Ceftiofur is an extended-spectrum cephalosporin licensed for the treatment of respiratory infections in pigs and cattle. Various authors have hypothesized that the veterinary use of extended-spectrum cephalosporins may select for extended-spectrum β-lactamase (ESBL)-producing Enterobacteriaceae in animals, resulting in an increased risk of the zoonotic transmission of ESBL-carrying bacteria and plasmids (2, 10, 11, 17, 18).. A statistical association between the prophylactic use of ceftiofur and the occurrence of cefotaxime-resistant E. coli in healthy pigs was recently demonstrated at two Danish pig farms, and most of the isolates were shown to be CTX-M-1 producers (11). In this study, we revisited the farms 1 year after the previous study to investigate the distribution, persistence, and transmission of blaCTX-M-1 between pigs, farm workers, and ...
Ribosomal Protein L39 CRISPR Plasmids | SCBT - Santa Cruz Biotechnology
산타크루즈바이오테크놀러지는 광범위한 유전자편집 제품을 제공하고 있으며 유전자침묵에 쓰이는 CRISPR/Cas9 Knockout 와 CRISPR Double Nickase plasmids를 제공합니다. Ribosomal Protein L39유전자침묵에는 Ribosomal Protein L39 CRISPR/Cas9 Knockout plasmids 와 Ribosomal Protein L39 Double Nickase Plasmids를 제공합니다. 또한 유전자활성화에 쓰이는 Ribosomal Protein L39 CRISPR/dCas9 Activation Plasmids와 CRISPR Lenti Activation Systems도 제공합니다. 유전자침묵과 활성화는 유전자연구에 유용하게 쓰이며 이는 항체와 결합하여 단백질의 검출에 유용하게 쓰입니다.
Ribosomal Protein L39L CRISPR Plasmids | SCBT - Santa Cruz Biotechnology
산타크루즈바이오테크놀러지는 광범위한 유전자편집 제품을 제공하고 있으며 유전자침묵에 쓰이는 CRISPR/Cas9 Knockout 와 CRISPR Double Nickase plasmids를 제공합니다. Ribosomal Protein L39L유전자침묵에는 Ribosomal Protein L39L CRISPR/Cas9 Knockout plasmids 와 Ribosomal Protein L39L Double Nickase Plasmids를 제공합니다. 또한 유전자활성화에 쓰이는 Ribosomal Protein L39L CRISPR/dCas9 Activation Plasmids와 CRISPR Lenti Activation Systems도 제공합니다. 유전자침묵과 활성화는 유전자연구에 유용하게 쓰이며 이는 항체와 결합하여 단백질의 검출에 유용하게 쓰입니다.
Vol 9: Complete Nucleotide Sequence and Analysis of Two Conjugative Broad Host Range Plasmids from a Marine Microbial Biofilm. ...
Vol 9: Complete Nucleotide Sequence and Analysis of Two Conjugative Broad Host Range Plasmids from a Marine Microbial Biofilm.. This article is from PLoS ONE, volume 9.AbstractThe complete nucleotide sequence of plasmids pMCBF1 and pMCBF6 was deter. Biblioteca virtual para leer y descargar libros, documentos, trabajos y tesis universitarias en PDF. Material universiario, documentación y tareas realizadas por universitarios en nuestra biblioteca. Para descargar gratis y para leer online.
APPENDIX B MAINTENANCE ALLOCATION CHART
TM 10 6670-277 13&P APPENDIX B MAINTENANCE ALLOCATION CHART Section I. INTRODUCTION B 1. General. a. This section provides a general explanation of all maintenance and repair functions authorized at various maintenance categories. b. The Maintenance Allocation Chart (MAC) in Section II designates overall authority and responsibility for the performance of maintenance functions on the identified end item or component. The application of the maintenance functions to the end item or component will be consistent with the capacities and capabilities of the designated maintenance categories. c. Section Ill lists the tools and test equipment (both special tools and common tool sets) required for each maintenance function as referenced from Section Il. d. Section IV contains supplemental instructions and explanatory notes for a particular maintenance function. B 2. Maintenance Functions. Maintenance functions will be limited to and defined as follows: a. Inspect. To determine the serviceability of an ...
High levels of foreign gene expression in hepatocytes after tail vein injections of naked plasmid DNA
We have previously shown that the intramuscular injection of naked plasmid DNA enables foreign gene expression in muscle. Further studies showed that the intravascular delivery of naked plasmid DNA enables high levels of expression not only in muscle but also in hepatocytes. For the liver, this tech …
Evolutionary Trajectory of the Replication Mode of Bacterial Replicons | mBio
Bacterial circular chromosomes replicate bidirectionally by the theta mechanism. Circular plasmids replicate by three general mechanisms: theta type, strand displacement, and rolling circle (43). Different machineries are involved in different plasmid replication mechanisms (43). For example, initiation of theta-type plasmid replication generally requires a plasmid-encoded initiator protein. In contrast, initiation of strand displacement replication requires the plasmid-encoded helicase, primase, and initiator protein, and initiation of rolling-circle replication requires the plasmid-encoded protein with DNA strand transferase enzymatic activity. The absence of the above essential proteins for the latter two mechanisms indicated that Pseudoalteromonas chromids replicate by the theta mechanism. Consistent with this, no such proteins in chromids from other bacteria have been reported. Considering the different replication factors involved in different mechanisms, a parsimony assumption is that ...
Evaluation of interspecies plasmid transfer and comparison with hospital epidemiology data | NIH Research Festival
Objective: Carbapenem-producing organisms have spread worldwide and cause significant morbidity. Horizontal gene transfer of carbapenemases may play a role in this spread. Given that conjugation is influenced by a number of factors, we sought to perform a systematic analysis of blaKPC encoding plasmid transfer into multiple species using hospital isolates. Methods: Plasmids were isolated from patient donor strains from the NIH and University of Virginia and a subset were tagged with GFP and electroporated into a K. pneumoniae patient isolate cured of its blaKPC plasmid. Broth and filter matings were performed, and transconjugants were isolated on selective media. Plasmids tested included those found in multiple species during hospital surveillance. Results: Transfer frequency was dependent on the recipient, temperature, substratum, and the specific plasmid. pKPC-47e was extremely attenuated in conjugation efficiency across all conditions tested compared to pKpQIL. In vitro studies showed a low ...
Publication : USDA ARS
Interpretive Summary: A novel plasmid (pHoss1) for Listeria monocytogenes was constructed. pHoss1 is very efficient for the generation of in-frame deletion mutants. The L. monocytogenes ispG and ispH genes were in-frame mutated. The L. monocytogenes ispG and ispH genes are not involved in cell adhesion. Technical Abstract: Although several plasmids have been used in Listeria monocytogenes for generating mutants by allelic exchange, construction of L. monocytogenes mutants has been inefficient due to lack of effective selection markers for first and second recombination events. To address this problem, we have developed a new suicide plasmid, pHoss1, by using the pMAD plasmid backbone and anhydrotetracycline selection marker (secY antisense RNA) driven by an inducible Pxyl/tetO promoter. Expression of the secY antisense RNA eliminates merodiploids and selects for the loss of plasmid via a second allelic exchange, which enriches the number of mutants with deleted genes. To assess the effectiveness ...
In the PBE lab we study the role of plasmids as catalysts of bacterial evolution, with a special focus on the evolution of plasmid-mediated antibiotic resistance.. Currently, we have two ongoing projects:. - In vivo evolution of plasmid-mediated resistance.. Conjugative plasmids play a key role in the horizontal spread of antibiotic resistance mechanisms among bacteria. One of the key factors undermining the successful spread of a conjugative plasmid is the initial fitness cost produced by the plasmid in the recipient bacteria. The factors involved in this cost and its potential compensation remain largely unknown. In our lab we are trying to understand the evolutionary and genetic determinants that promote the emergence and establishment of successful associations between bacterial clones and resistance plasmids in vivo.. To do so we study conjugation events between different enterobacteria occurring in the gut of hospitalized patients. We study the cost produced by the plasmids when they first ...
Restriction site free cloning (RSFC) plasmid family for seamless, sequence independent cloning in Pichia pastoris | Microbial...
Tagging proteins is a standard method facilitating protein detection, purification or targeting. When tagging a certain protein of interest, it is challenging to predict which tag will give optimal results and will not interfere with protein folding, activity or yields. Ideally, multiple tags and positions are tested which however complicates molecular cloning and expression vector generation. In conventional cloning, tags are either added on PCR primers (requiring a distinct primer and PCR product per tag) or provided on the vector (typically leaving a restriction site scar). Here we report a vector family of 40 plasmids allowing simple, seamless fusions of a single PCR product with various N- and C-terminal tags, signal sequences and promoters. The restriction site free cloning (RSFC) strategy presented in this paper relies on seamless cloning using type IIS restriction endonucleases. After cutting out a stuffer (placeholder) fragment from the vectors, a single PCR product can be directly inserted in
Difference between revisions of Synthetic Biology:Vectors/Single copy plasmid - OpenWetWare
The F plasmid origin needs to be designed. The complete F plasmid with partitioning genes in ~10kb in length. It contains several BioBricks restriction sites in both coding and noncoding regions. Once designed, the F plasmid origin can be assembled with an antibiotic resistance marker and cloned into the vector scaffold to generate a new single copy BioBricks plasmid. Chris Anderson suggested inclusion of the R6K origin in these plasmids (rather than inclusion of a pUC19 origin in the multiple cloning site). The R6K origin is a conditional origin. It only works in the presence of the trans-acting protein Π (encoded by pir) for replication. R6K replicates at a medium copy (15 per cell) in pir+ strains and high copy (250 per cell) in pir-116 (high-copy-number mutant) E. coli hosts. ...
Difference between revisions of Synthetic Biology:Vectors/Single copy plasmid - OpenWetWare
The F plasmid origin needs to be designed. The complete F plasmid with partitioning genes in ~10kb in length. It contains several BioBricks restriction sites in both coding and noncoding regions. Once designed, the F plasmid origin can be assembled with an antibiotic resistance marker and cloned into the vector scaffold to generate a new single copy BioBricks plasmid. Chris Anderson suggested inclusion of the R6K origin in these plasmids (rather than inclusion of a pUC19 origin in the multiple cloning site). The R6K origin is a conditional origin. It only works in the presence of the trans-acting protein Π (encoded by pir) for replication. R6K replicates at a medium copy (15 per cell) in pir+ strains and high copy (250 per cell) in pir-116 (high-copy-number mutant) E. coli hosts. ...
Gene doctoring: a method for recombineering in laboratory and pathogenic Escherichia coli strains - ePrints Repository
Results: Our goal was to develop a high-throughput recombineering system, primarily for the coupling of genes to epitope tags, which could also be used for deletion of genes in both pathogenic and K-12 E. coli strains. To that end we have designed a series of donor plasmids for use with the lambda-Red recombination system, which when cleaved in vivo by the I-SceI meganuclease generate a discrete linear DNA fragment, allowing for C-terminal tagging of chromosomal genes with a 6xHis, 3xFLAG, 4xProteinA or GFP tag or for the deletion of chromosomal regions. We have enhanced existing protocols and technologies by inclusion of a cassette conferring kanamycin resistance and, crucially, by including the sacB gene on the donor plasmid, so that all but true recombinants are counter-selected on kanamycin and sucrose containing media, thus eliminating the need for extensive screening. This method has the added advantage of limiting the exposure of cells to the potential damaging effects of the lambda-Red ...
Vaccination: Plasmid DNA Vaccines | GreenMedInfo | Anti-Therapeutic
This topic contains 2 study abstracts on Vaccination: Plasmid DNA Vaccines indicating they may negatively impact Infertility, Vaccination: Abortion, and Vaccine-induced Toxicity
pCMV-GLuc 2 Control Plasmid | NEB
The pCMV-GLuc 2 Control Plasmid is a mammalian expression vector that encodes the secreted luciferase from the copepod Gaussia princeps as a reporter, under the control of the constitutive CMV (cytomegalovirus) promoter. Gaussia luciferase (GLuc) is a 19 kDa protein encoded by a humanized sequence, and it contains a native signal peptide at the N-terminus that allows it to be secreted from mammalian cells into the cell culture medium. A neomycin resistance gene under the control of an SV40 promoter allows selection for stable integration of the plasmid into the mammalian cell genome using G418.
Plasmids can stably transform yeast mitochondria lacking endogenous mtDNA | PNAS
The mitochondrial gene oxi1, carried on a bacterial plasmid, has been used to transform the mitochondria of a yeast strain lacking mtDNA (rho0). The plasmid DNA behaved in a manner entirely consistent with the known properties of normal yeast rho- mtDNA after its introduction by high-velocity microprojectile bombardment. Like the mtDNA sequences retained in natural rho- strains, the plasmid DNA in the transformants was reiterated into concatemers whose size was indistinguishable from that of wild-type mtDNA. The oxi1 sequences in the transformants were surrounded by restriction sites derived from the plasmid that were not present in wild-type mtDNA. oxi1 genetic information in these synthetic rho- strains could be expressed in diploids either after marker rescue by recombination with rho+ mtDNA carrying an appropriate oxi1 point mutation or in trans during the growth of diploids heteroplasmic for both the plasmid-derived oxi1 sequences and rho+ mtDNA with oxi1 deleted. The ability to ...
Help: 2 Plasmid Electroporation /Infection
I was hoping for suggestions as I am having difficulty getting two plasmids into E.coli at library efficiences. I am hoping to add a M13 derived vector to cells containing a pUC/pBR3222 plasmid, or vice versa. Electroporating the M13 derived vector has been at low efficiency so far, making double electroporations very inefficient. The cells containing these plasmids were less electrocompetent, less than 10^6 col/ ug of the second plasmid. Therefore, this approach would require many electroporations to get a large library. I next looked to make the M13 derived vector infectious by growing with helper phage, but had too much cross over with the helper phage. I am now trying this in a Rec minus strain. I was hoping to electroporate in one plasmid and then add the other plasmid through lambda infection but was unable to locate a non lysing system. Please let me know if you have had similar experiences or any suggestions. Thanks tj t j cradick ucsf dept of immunology please reply to:tjc at ...
Isolation of plasmid DNA from Bacillus subtilis using the QIAGEN Plasmid Midi Kit
The procedure has been used successfully for isolation of high- and low-copy-number plasmids from various Bacillus subtilis strains. Yield of plasmid DNA was typically 10-20 µg plasmid DNA from 100 ml culture ...
In the PBE lab we are interested in the evolutionary forces that drive plasmid dynamics in bacterial populations. Plasmids play a crucial role in bacterial ecology and evolution because they can transfer genes horizontally between different bacteria. The most striking example of how plasmids drive bacterial evolution is the global spread of plasmid-mediated antibiotic resistance over the last few decades. Plasmids are arguably the main vehicle for the spread of antibiotic resistance genes among clinically relevant bacteria, contributing to the overwhelming antibiotic resistance crisis we are currently facing.. In our group we try to understand the population genetics of antibiotic resistance plasmids using advanced molecular and evolutionary techniques. Ultimately, we intend to apply the concepts that we learn from the study of the evolution of plasmid-mediated antibiotic resistance to develop more rational intervention strategies to control infectious diseases.. ...
Complete Sequence of pSAM7, an IncX4 Plasmid Carrying a Novel bla[sub]CTX-M-14b Transposition Unit Isolated from Escherichia...
The same plasmid carrying blaCTX-M-14b was identified from an Escherichia coli isolate and an Enterobacter cloacae isolate collected from cattle in the United Kingdom by complete plasmid sequencing. This 35,341-bp plasmid, pSAM7, had an IncX4 backbone that is 99% identical to that of pJIE143 from a human isolate in Australia. PCR screening identified pSAM7-like plasmids in three other E. coli isolates of different multilocus sequence types isolated from cattle on different farms in the United Kingdom.. ...
Characterization of a Theta-Type Plasmid from Lactobacillus sakei: a Potential Basis for Low-Copy-Number Vectors in...
pRV500 is, to our knowledge, the first plasmid of L. sakei to be entirely sequenced. Careful sequence analysis showed that pRV500 belongs to the pUCL287 family, itself related to class A theta-type plasmids of the pUCL22 and pSC101 families. Experimental evidence of the theta replication mode was obtained for pUCL287 itself (2). Although the minimum length of the pRV500 oriA region remains to be determined, it is likely to involve the same two kinds of DNA repeats as other pUCL287-type plasmids. The involvement of repetitive sequences of 22 bp (iterons) as a target for binding of the Rep protein was demonstrated for pSBO1 (40), and the upstream 11-bp repeated sequences were observed to be essential for pUCL287 replication (3).. Like other plasmids (14, 49), pRV500 appears to be a composite structure containing DNA segments from different sources. Some ORFs are shared by plasmids from both subfamilies pUCL22 and pUCL287; this applies to orf6 in pRV500 and ORFs of unknown function around the gene ...
Isolation of indigenous wastewater bacterial strains capable of mobilizing plasmid pBR325. | Applied and Environmental...
Members of the family Enterobacteriaceae have been isolated from raw wastewater, identified, and characterized with respect to their plasmid content and antibiotic resistance. Several strains possessing both antibiotic resistance and high-molecular-weight plasmid(s) transferred their resistance characteristics to recipient cells during a 25 h coincubation. Eight were characterized (six Escherichia coli and two Klebsiella pneumoniae); each produced 10(2) to 10(7) transconjugants per ml by the end of the incubation period. They were also able to mobilize pBR325 from a laboratory E. coli strain into plasmid-free recipients to yield 10(2) to 10(7) transconjugants per ml. These transconjugants possessed phenotypic characteristics specified by pBR325, the R plasmid, and the chromosome of the recipient. Many transconjugants exhibited recombinational rearrangements of the acquired plasmid material. ...
Replicon Typing of Plasmids Encoding Resistance to Newer β-Lactams - Volume 12, Number 7-July 2006 - Emerging Infectious...
Polymerase chain reaction-based replicon typing represents a novel method to describe the dissemination and follow the evolution of resistance plasmids. We used this approach to study 26 epidemiologically unrelated Enterobacteriaceae and demonstrate the dominance of incompatibility (Inc) A/C or Inc N-related plasmids carrying some emerging resistance determinants to extended-spectrum cephalosporins and carbapenems.
Expression Plasmids | Bioz
Bioz Stars score, Techniques, Protocol Conditions and more for Expression Plasmids, supplied by . Data for Expression Plasmids gathered from related PubMed articles.
Detection of conjugative plasmids and evaluation of genotoxicity
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Book The Biology Of Plasmids
book The Biology of Plasmids is ordered for your wall. Some stakeholders of this book The Biology of Plasmids may together vary without it. Oleg ShmygulLoading PreviewSorry, book The motivates fully other.
SPINACH PLASMIDS. ...
Biotechnology in pharmaceutical manufacturing
Plasmids containing the Factor IX gene, along with plasmids with a gene that codes for resistance to methotrexate, were ... The recombinant plasmids were inserted into Escherichia coli bacteria, which were "induced to produce 100,000 molecules of ... The gene that was inserted into the plasmid was created by reverse transcription of the mRNA found in pituitary glands to ... The artificial genes were "then inserted... into plasmids... among a group of genes that" are activated by lactose. Thus, the ...
It is used to increase the transfection efficiency of RNA (including mRNA and siRNA) or plasmid DNA into in vitro cell cultures ... DNA plasmids. Lipofectamine consists of a 3:1 mixture of DOSPA (2,3‐dioleoyloxy‐N‐ [2(sperminecarboxamido)ethyl]‐N,N‐dimethyl‐1 ...
Fertility plasmids, or f plasmids, allow for conjugation to occur whereas resistance plasmids, or r plasmids, contain genes ... Circular bacterial plasmids are classified according to the special functions that the genes encoded on the plasmid provide. ... Circular bacterial plasmids are also the basis for the production of DNA vaccines. Plasmid DNA vaccines are genetically ... The linear plasmids with a covalently attached protein may assist with bacterial conjugation and integration of the plasmids ...
Plasmids in Bacteria. Boston, MA: Springer US. ISBN 1-4613-2447-5.CS1 maint: extra text: authors list (link) Roberts, edited by ...
Plasmid. 61 (3): 171-175. doi:10.1016/j.plasmid.2008.12.002. PMC 2699751. PMID 19167423. Labes, G.; Bibb, M.; Wohlleben, W. (1 ... Plasmid. 61 (3): 171-175. doi:10.1016/j.plasmid.2008.12.002. PMC 2699751. PMID 19167423. ATCC Type strain of Streptomyces ... Muth, Gunter; Wohlleben, Wolfgang; Pohler, Alfred (March 1988). "The minimal replicon of the Streptomyces ghanaensis plasmid ... Plasmids in Bacteria. Boston, MA: Springer US. ISBN 1-4613-2447-5. George M. Garrity, ed. (2012). Bergey's manual of systematic ...
As of 2014 Addgene's repository comprised 30,000 plasmids, deposited by 1,700 labs. Its plasmid collection contains plasmids ... Plasmid Cloning Guides Molecular Cloning Guides-References to help scientists design plasmid cloning experiments, including ... The plasmids are available to both academic and industry labs. Noteworthy depositors include: 12 Nobel Prize ... Addgene accepts plasmids from researchers for distribution and archival. Addgene obtains revenues and licensing fees using the ...
Plasmids in Bacteria. Boston, MA: Springer US. ISBN 978-1-4613-2447-8.CS1 maint: extra text: authors list (link) I.W. Southon; ...
Mobile genetic elements
David Summers (1996). "Chapter 1 - The Function and Organization of Plasmids". The Biology of Plasmids (First ed.). Wiley- ... Plasmids during their cycle carry genes from one organism to another through a process called conjugation. They also often ... MGEs such as plasmids by a horizontal transmission are generally beneficial to an organism. The ability of transferring ... Plasmids: They are generally circular extrachromosomal DNA molecules that replicate and are transmitted independent of ...
Recombinant AAV mediated genome engineering
Kandavelou K, Chandrasegaran S (2008). "Plasmids for Gene Therapy". In Lipps, Georg (ed.). Plasmids: Current Research and ...
Zinc finger nuclease
Kandavelou K, Chandrasegaran S (2008). "Plasmids for Gene Therapy". Plasmids: Current Research and Future Trends. Norfolk: ... Since ZFN-encoding plasmids could be used to transiently express ZFNs to target a DSB to a specific gene locus in human cells, ... The ZFN-encoding plasmid-based approach has the potential to circumvent all the problems associated with the viral delivery of ... The construction of plasmid vectors is simple and straightforward. Custom-designed ZFNs that combine the non-specific cleavage ...
The two plasmids are pTiC58, responsible for the processes involved in virulence, and pAtC58, dubbed the "cryptic" plasmid. The ... To be virulent, the bacterium contains tumour-inducing plasmid (Ti plasmid or pTi), of 200 kbp, which contains the T-DNA and ... tumefaciens infects the plant through its Ti plasmid. The Ti plasmid integrates a segment of its DNA, known as T-DNA, into the ... "The cryptic plasmid of Agrobacterium tumefaciens cointegrates with the Ti plasmid and cooperates for opine degradation". ...
Maia M.; Sanchez J. M.; Vela G. R. (1988). "Plasmids of Azotobacter vinelandii". Journal of Bacteriology. 170 (4): 1984-1985. ... In addition to chromosomal DNA, Azotobacter can contain plasmids. Azotobacter species are ubiquitous in neutral and weakly ...
Many of the weapons and plasmids from BioShock return, as well as new ones like a chain gun and the Aero Dash plasmid. Players ... Upgraded versions of plasmids bestow additional abilities for more tactical plasmid usage; for example, an upgraded Electro ... The citizen is engaged by the plasmid manufacturer Sinclair Solutions to test out their weapons and plasmids as part of a ... and plasmids. Plasmids function akin to magic and must be recharged with "EVE" to be used. The player can also equip "tonics", ...
Enterobacteria phage P4
The P4 genome can also exist on its own within the host cell and can replicate as a free plasmid. Pruss, G; Goldstein, RN; ... Briani, Federica; Dehò, Gianni; Forti, Francesca; Ghisotti, Daniela (2002). "The Plasmid Status of Satellite Bacteriophage P4 ... ". Plasmid. 45 (1): 1-17. doi:10.1006/plas.2000.1497. PMID 11319927. Claverie JM, Abergel C (2009). "Mimivirus and its ...
Kolter, R; Helinski, DR (1982). "Plasmid R6K DNA replication. II. Direct nucleotide sequence repeats are required for an active ... Kolter, R; Helinski, DR (1978). "Construction of plasmid R6K derivatives in vitro: characterization of the R6K replication ... "Trans-complementation-dependent replication of a low molecular weight origin fragment from plasmid R6K". Cell. 15 (4): 1199-208 ... region". Plasmid. 1 (4): 571-80. doi:10.1016/0147-619X(78)90014-8. PMID 372982. Kolter, R; Inuzuka, M; Helinski, DR (Dec 1978 ...
Cleavage of a plasmid in Halobacterium halobium resulted in the loss of the ability to biosynthesize gas vesicles, indicating ... DasSarma S, Damerval T, Jones JG, Tandeau de Marsac N (November 1987). "A plasmid-encoded gas vesicle protein gene in a ... Weidinger G, Klotz G, Goebel W (July 1979). "A large plasmid from Halobacterium halobium carrying genetic information for gas ... Although there is evidence suggesting the early evolution of gas vesicles, plasmid transfer serves as an alternate explanation ...
... faecalis plasmid pCF10 conjugation". Plasmid. 64 (1): 26-35. doi:10.1016/j.plasmid.2010.03.002. PMC 2892192. PMID 20332003. ... Anti-Q RNA (formerly Qa RNA) is a small ncRNA from the conjugal plasmid pCF10 of Enterococcus faecalis. It is coded in cis to ... Brantl S, Birch-Hirschfeld E, Behnke D (1993). "RepR protein expression on plasmid pIP501 is controlled by an antisense RNA- ... Novick RP, Iordanescu S, Projan SJ, Kornblum J, Edelman I (1989). "pT181 plasmid replication is regulated by a ...
Multicopy single-stranded DNA
Plasmid. 33 (3): 235-8. doi:10.1006/plas.1995.1026. PMID 7568472. Hsu MY, Inouye M, Inouye S (December 1990). "Retron for the ...
For example, in plasmid ColE1, the asRNA termed RNA I plays an important role in determining the plasmid copy number by ... Plasmid. 114: 102560. doi:10.1016/j.plasmid.2021.102560. PMID 33482228. "alpha thalassemia". Genetics Home Reference. NIH U.S. ... The initial asRNAs discovered were in prokaryotes including plasmids, bacteriophage and bacteria. ...
Nabeshima, S; Hotta, Y; Okanishi, M (September 1984). "Construction of plasmid vectors from Streptomyces kasugaensis plasmids, ... Toyama, Hiromi; Okanishi, Masanori; Umezawa, Hamao (May 1981). "Physical characterization of plasmids from Streptomyces ... kasugaensis MB273". Plasmid. 5 (3): 306-312. doi:10.1016/0147-619X(81)90007-X. PMID 6267633. Chae, Won-Bok; Kim, Young-Bum; ...
Plasmids often carry genes that are responsible for bacterial antibiotic resistance; as these plasmids replicate and pass from ... the most common mobile genetic elements in the prokaryotic genome are plasmids and prophages. Plasmids and prophages can move ... Plasmids were discovered in the 1940s as genetic materials outside of bacterial chromosomes. Prophages are genomes of ... Toussaint A, Merlin C (January 2002). "Mobile elements as a combination of functional modules". Plasmid. 47 (1): 26-35. doi: ...
Plasmid. 110 (102505): 102505. doi:10.1016/j.plasmid.2020.102505. PMID 32380021. Type strain of Acinetobacter lwoffii at ...
Plasmid-borne streptothricin resistance in gram-negative bacteria. Plasmid. 12:3 189-196. Miyadoh S. 1993. Research on ... pIJ101, a multi-copy broad host-range Streptomyces plasmid: functional analysis and development of DNA cloning vectors. Mol Gen ... One study found genes for streptothricin resistance, an antibiotic produced by Actinomycete bacteria, on plasmids within gram- ... In addition, actinomycete genomes contain extrachromosomal genetic elements such as rolling circle replication plasmids. These ...
Origin of replication
It further rationalizes plasmid incompatibilities in E. coli, where certain plasmids destabilize each other's inheritance due ... Brewer BJ, Fangman WL (November 1987). "The localization of replication origins on ARS plasmids in S. cerevisiae". Cell. 51 (3 ... Novick RP (December 1987). "Plasmid incompatibility". Microbiological Reviews. 51 (4): 381-95. doi:10.1128/MMBR.51.4.381- ... Plasmid. 23 (2): 159-62. doi:10.1016/0147-619x(90)90036-c. PMID 2194231. Paululat A, Heinisch JJ (December 2012). "New yeast/E ...
It was first found on plasmid pNDH33. The plasmid also carries replication regions from the pMTLBs72. The plasmid also carries ... 2003). "Bacillus subtilis soil isolates: plasmid replicon analysis and construction of a new theta-replicating vector". Plasmid ... pHT01 is a plasmid used as a cloning vector for expressing proteins in Bacillus subtilis. It is 7,956 base pairs in length. ... Plasmid pHT01 is generally stable in both B. subtilis and Escherichia coli, and can be used for protein expression in these ...
Plasmid. 31 (2): 222-8. doi:10.1006/plas.1994.1024. PMID 8029330. Mazauric MH, Licznar P, Prère MF, Canal I, Fayet O (July 2008 ...
... a multiple cryptic plasmid containing strain". Plasmid. 65 (2): 110-117. doi:10.1016/j.plasmid.2010.11.003. PMID 21087624. Type ... "Physiological and biochemical consequences of host-plasmid interaction - A case study with Corynebacterium renale, ...
Larson, J.L.; Hershberger, C.L. (May 1990). "Insertion of plasmids into the chromosome of Streptomyces griseofuscus". Plasmid. ...
Plasmids have been found to not function in S. cellulosum cells. Reproducible genetic alterations must be made directly into ... Jaoua, S.; Neff, S.; Schupp, T. (1992). "Transfer of mobilizable plasmids to Sorangium cellulosum and evidence for their ... integration into the chromosome". Plasmid. 28 (2): 157-165. doi:10.1016/0147-619x(92)90046-d. PMID 1409972. Lee FY, Borzilleri ...
Plasmid RNAIII - Wikipedia
Plasmid preparation - Wikipedia
A typical plasmid DNA yield of a miniprep is 5 to 50 µg depending on the cell strain. Miniprep of a large number of plasmids ... Minipreparation of plasmid DNA is a rapid, small-scale isolation of plasmid DNA from bacteria. It is based on the alkaline ... A plasmid preparation is a method of DNA extraction and purification for plasmid DNA. Many methods have been developed to ... The plasmid DNA yield will vary depending on the plasmid copy number, type and size, the bacterial strain, the growth ...
Conjugative Plasmids of Streptomyces | SpringerLink
Terminal Inverted Repeat Linear Plasmid Plasmid Transfer Mycobacterium Smegmatis Conjugative Plasmid These keywords were added ... Holloway, B. W., 1979, Plasmids that mobilize bacterial chromosome, Plasmid 2: 1-19.PubMedCrossRefGoogle Scholar ... Hopwood, D. A., Lydiate, D. J., Malpartida, F., and Wright, H. M., 1985, Conjugative plasmids in Streptomyces, in: Plasmids in ... a conjugative plasmid will mobilize nontransferable plasmids in soil, Curr. Microbiol. 19: 115-121.Google Scholar ...
What are Plasmids?
... plasmids. They can also be present at much lower frequencies in certain eukaryotic cell types, such as yeast. They are non- ... These plasmids can exist in two forms, which are extrachromosomal replicons or integrated plasmids. These plasmids are known as ... have a unique location in that plasmid. These sites flank the cloning site in the plasmid. Many modified plasmids have a ... This makes the plasmid a more flexible cloning vector.. Modified plasmids also carry a marker gene. The most common form is a ...
Plasmid-mediated resistance - Wikipedia
Plasmid-mediated resistance is the transfer of antibiotic resistance genes which are carried on plasmids. The plasmids can be ... Other smaller plasmids (usually < 10 kb in size) can be mobilized by a conjugative plasmid (usually > 30 kb) in order to be ... are common for resistance plasmids in Enterobacteriaceae. Often multiple beta-lactamase genes are found on the same plasmid ... It is very common for the resistance genes or entire resistance cassettes to be re-arranged on the same plasmid or be moved to ...
Definitions of plasmids - OneLook Dictionary Search
plasmids: Vocabulary.com [home, info] *Plasmids, plasmids: Wordnik [home, info] *plasmids: Wiktionary [home, info] *plasmids: ... Phrases that include plasmids: pet plasmids Words similar to plasmids: plasmid, more... ... plasmids: Urban Dictionary [home, info] (Note: See plasmid for more definitions.). Quick definitions from WordNet (plasmid). ▸ ... Plasmids: Encyclopedia [home, info] Medicine (4 matching dictionaries). *Plasmids: Medical Dictionary [home, info] *plasmids: ...
... everything you need for studying or teaching Plasmid. ... Immediately download the Plasmid summary, chapter-by-chapter ... Plasmid Plasmids are naturally occurring, stable genetic elements found in bacteria, fungi, and even in the mitochondria of ... Plasmids Plasmids are extra-chromosomal, covalently closed circular (CCC) molecules of double stranded (ds) DNA that are ... Plasmids Plasmids are extra-chromosomal, covalently closed circular (CCC) molecules of double stranded (ds) DNA that are ...
... Kevin ODonnell odonnell at sasa.gov.uk Wed Dec 13 12:22:28 EST 1995 *Previous message: TOL plasmids ... the presence of such a plasmid. Does anybody have a protocol that will ,let me visualize such a plasmid on a gel? ( ... In article ,4ai9os$up at news2.ucsd.edu,, rcaspi at sdcc3.ucsd.edu (Ron Caspi) says: , ,Hi, is there a TOL plasmid expert out ... unfortunately, pulse field ,is not available). , ,Many thanks in advance , I did my PhD on a TOL plasmid from Pseudomonas ...
Plasmid | microbiology | Britannica.com
Plasmids are circular deoxyribonucleic acid (DNA) molecules that replicate independently of the bacterial chromosome. They are ... Plasmid, in microbiology, an extrachromosomal genetic element that occurs in many bacterial strains. ... Plasmid, in microbiology, an extrachromosomal genetic element that occurs in many bacterial strains. Plasmids are circular ... small extra DNA molecules called plasmids. Bacterial plasmids are circular DNA molecules; some carry genes for resistance to ...
Plasmids + copies of the DNA fragment produce quantities of recombinant DNA.. This recombinant DNA stew is allowed to transform ... The plasmid is opened up and the gene is freed from its parent DNA strand. They have complementary sticky ends. The opened ... In DNA cloning, a DNA fragment that contains a gene of interest is inserted into a cloning vector or plasmid. ... The plasmid carrying genes for antibiotic resistance, and a DNA strand, which contains the gene of interest, are both cut with ...
Planning Plasmids | Science
Antibiotic Resistance-Carrying Plasmids [image] | EurekAlert! Science News
The result is plasmids consisting of genes that have each been adapted to different bacterial species. This facilitates further ... Antibiotic resistance-carrying plasmids from different bacteria can meet and exchange genetic material. ... Antibiotic resistance-carrying plasmids from different bacteria can meet and exchange genetic material. The result is plasmids ...
E. coli Plasmid Vectors | SpringerLink
Comprehensive and highly practical, E. coli Plasmid Vectors offers those new to the field a basic guide to the use of plasmid ... The authors describe readily reproducible methods for cloning DNA into plasmid vectors, transforming plasmids into E. coli, and ... In E. coli Plasmid Vectors, experienced bench researchers describe their proven techniques for the manipulation of recombinant ...
Plasmids - dummies
Aside from conjugative plasmids, the other main types of plasmids are. * Resistance plasmids: Carry genes for resistance to ... theyre called conjugative plasmids. Conjugative plasmids can facilitate the transfer of themselves, other plasmids, and even ... Some plasmids exist in many copies inside one cell and are called high-copy-number plasmids, whereas others are less numerous ... Although the enzymes involved in DNA replication of plasmids are the same as those used for the chromosome, some plasmids are ...
KorSA (plasmid) [Streptomyces ambofaciens] - Protein - NCBI
Cloning Plasmids and DNAs | NEB
Cloning Plasmid/DNA NEB # Features Concentration MW/Size Size Price pBR322 Vector N3033S/L Commonly used cloning vectors Tet ... Home Tools & Resources Selection Charts Cloning Plasmids and DNAs Cloning Plasmids and DNAs. Cloning Plasmid/DNA. NEB #. ... Kits are available for total RNA purification, plasmid miniprep, gel extraction, and DNA & RNA cleanup. For maximum convenience ... Kits are available for total RNA purification, plasmid miniprep, gel extraction, and DNA & RNA cleanup. For maximum convenience ...
... Dominic-Luc Webb molmed domweb at mbox.ki.se Fri Jul 13 03:43:17 EST 2001 *Previous message: Shine Dalgano ... Does anyone know of a database with plasmid sequences? Alternatively, I would like to know vendor for pCMX plasmid. Cheers, ... I am trying to find the sequence of some plasmids, but I have no idea who the vendor is, etc... ...
Plasmids can stably transform yeast mitochondria lacking endogenous mtDNA | PNAS
Plasmids can stably transform yeast mitochondria lacking endogenous mtDNA. T D Fox, J C Sanford, and T W McMullin ... Plasmids can stably transform yeast mitochondria lacking endogenous mtDNA Message Subject (Your Name) has sent you a message ... The mitochondrial gene oxi1, carried on a bacterial plasmid, has been used to transform the mitochondria of a yeast strain ... The oxi1 sequences in the transformants were surrounded by restriction sites derived from the plasmid that were not present in ...
Studies on transformation of Escherichia coli with plasmids. - PubMed - NCBI
Transformation efficiency declines linearly with increasing plasmid size. Relaxed and supercoiled plasmids transform with ... Studies on transformation of Escherichia coli with plasmids.. Hanahan D.. Abstract. Factors that affect the probability of ... A set of conditions is described under which about one in every 400 plasmid molecules produces a transformed cell. These ... Competition with both transforming and non-transforming plasmids indicates that each cell is capable of taking up many DNA ...
Antibiotic-resistant plasmids flourish in hospital plumbing | EurekAlert! Science News
They conducted whole-genome analyses on the samples to study the bacterial plasmids, or rings of DNA, that can confer ... Understanding the plasmid exchange and when the plasmids get into the pathogens that infect our patients, she says, could help ... Antibiotic-resistant plasmids flourish in hospital plumbing. American Society for Microbiology. Journal. mBio. Keywords. * ... "How much should we care that there are a bunch of plasmids down in the wastewater system if theyre not infecting our patients ...
Recent Articles | Plasmids | The Scientist Magazine®
Plasmid Partitioning | Science
plasmid drawing program
HiSpeed Plasmid Kits ( ...)
For ultrafast purification ... Principle ...,HiSpeed,Plasmid,Kits,biological,advanced biology technology,biology laboratory ... Basic Plasmid DNA Isolation Protocol. 9. RNA-free Plasmid DNA Isolation Protocol. 10. Basic Plasmid DNA Isolation Protocol. 11 ... Up to 200 g (Midi) or 750 g (Maxi) yield of high-copy plasmid DNA * Actual yields depend on plasmid copy number, size of insert ... New Reporter Plasmids for Studying Interferon-Stimulated Signal Transduction Pathways. 6. New Fusion Trans-Activator Plasmids ...
Plasmid - Journal - Elsevier
Plasmid publishes original research on genetic elements in all kingdoms of life with emphasis on maintenance, transmission and ... Plasmids promote the immigration of other plasmids but repress co-colonizing plasmids. ... Plasmids promote the immigration of other plasmids but repress co-colonizing plasmids. ... Recently published articles from Plasmid * Evolution in situ of ARI-A in pB2-1, a type 1 IncC plasmid recovered from Klebsiella ...
Patent US5981501 - Methods for encapsulating plasmids in lipid bilayers - Google Patents
The particles are typically 65-85 nm, fully encapsulate the plasmid and are serum-stable. ... Plasmid-lipid particles which are useful for transfection of cells in vitro or in vivo are described. The particles can be ... No plasmid was recovered in the absence of DODAC while, at a DODAC:plasmid charge ratio of 2:1, 90% of the plasmid was ... 10, the free plasmid eluted in the void volume of the column while, in FIG. 11, the plasmid incubated in serum eluted in the ...
Research shows new role for plasmids in antibiotic resistance
A new study led by scientists at the University of Oxford has found that small DNA molecules known as plasmids are one of the ... This occurs because bacteria usually carry more than one copy of a plasmid, which allows resistance genes carried by plasmids ... Our research shows a new role for plasmids in antibiotic resistance by demonstrating that plasmids drive the evolution of novel ... A new study led by scientists at the University of Oxford has found that small DNA molecules known as plasmids are one of the ...
Resistance plasmid | Define Resistance plasmid at Dictionary.com
What Is F Plasmid? | Reference.com
The F plasmid, also known as the fertility factor or sex factor, contains approximately 100 genes. Through conjugation, the F ... During conjugation, gene transfer between bacteria is driven by a DNA plasmid, such as the F plasmid, in the bacterial cells. ... The plasmid is a circular strand of DNA. Once transferred, it can replicate its own DNA and be transferred to other bacterial ... The F plasmid, also known as the fertility factor or sex factor, contains approximately 100 genes. Through conjugation, the F ...
CRACKING THE CODE/CLONING PAPER PLASMID
Ligate (scotch tape) the Vaccinia DNA and the Plasmid pBr322 vector. You NOW have a recombinant DNA plasmid that codes for the ... The vector DNA (plasmid) containing the "new" genes must now be inserted into host cells. The cells are made porous to DNA by a ... CRACKING THE CODE/CLONING PAPER PLASMID. Patricia Colbert and Phillis Unbehagen. 1993 Woodrow Wilson Biology Institute ... Isolate (cut out) the pBR322 DNA and circularize it into a small plasmid by using tape to connect the free ends. Be sure to ...
BacteriaReplicationGeneColiLinear plasmidsVectorsBacteriumConjugationDeoxyribonucleic acidIsolationGeneticVectorStrainsRestrictionChromosomeAntibiotic resistance genesMolecularLarger plasmidsExtra-chromosomalPurifyVirulencePresence of multiple plasmidsMoleculesChromosomalCarry genesConferGenomicAlkaline lysis methodCDNANonprofit plasmid repositoryProteinHigh-copy-number plasmidsRepliconsChromosomesMoleculeSingle-copy plasmidsEpisomesSite-Directed MuTransformationAnother plasmidSize of plasmidsWherein said plasmidCloningReplicateTransfectionAgarose gel analysisResistance plasmidPGLO plasmidLARGE PLASMIDSSmall plasmidsControl plasmidDifferent plasmidIdentical plasmidsRecombinant plasmidCopiesReagentsNaturally
- Many methods have been developed to purify plasmid DNA from bacteria. (wikipedia.org)
- These methods invariably involve three steps: Growth of the bacterial culture Harvesting and lysis of the bacteria Purification of plasmid DNA Plasmids are almost always purified from liquid bacteria cultures, usually E. coli, which have been transformed and isolated. (wikipedia.org)
- Virtually all plasmid vectors in common use encode one or more antibiotic resistance genes as a selectable marker, for example a gene encoding ampicillin or kanamycin resistance, which allows bacteria that have been successfully transformed to multiply uninhibited. (wikipedia.org)
- Bacteria that have not taken up the plasmid vector are assumed to lack the resistance gene, and thus only colonies representing successful transformations are expected to grow. (wikipedia.org)
- Minipreparation of plasmid DNA is a rapid, small-scale isolation of plasmid DNA from bacteria. (wikipedia.org)
- The plasmids can be transferred between bacteria within the same species or between different species via conjugation. (wikipedia.org)
- Antibiotic resistance mediated by MDR plasmids severely limits the treatment options for the infections caused by Gram-negative bacteria, especially Enterobacteriaceae family. (wikipedia.org)
- Plasmid Plasmids are naturally occurring, stable genetic elements found in bacteria, fungi, and even in the mitochondria of some plants. (bookrags.com)
- Keil et al, J.Bact vol. 163:248-255 The plasmids when you grow the bacteria up on nutrient broth prior to the sucrose gradient procedure - but growing them on benzoate seems to select all kinds of mutant phenotypes! (bio.net)
- One class of plasmids, colicinogenic (or Col ) factors, determines the production of proteins called colicins, which have antibiotic activity and can kill other bacteria. (britannica.com)
- Another class of plasmids, R factors, confers upon bacteria resistance to antibiotics. (britannica.com)
- Antibiotic resistance-carrying plasmids from different bacteria can meet and exchange genetic material. (eurekalert.org)
- Genetic information not contained in the chromosome of bacteria or archaea is kept as circular double-stranded DNA molecules called plasmids (although some linear plasmids do exist). (dummies.com)
- One important feature of plasmids is that they can be transferred between bacteria. (dummies.com)
- Bacteria die releasing their plasmids and other bacteria take them up. (dummies.com)
- Conjugative plasmids can facilitate the transfer of themselves, other plasmids, and even chromosomal DNA between bacteria. (dummies.com)
- The bacteria fight with each other and plasmids can carry genes that help them survive," she says. (eurekalert.org)
- Using a novel experimental model, the international team of researchers show that plasmids, which live inside bacteria and are known to be a vehicle for transferring antibiotic resistance genes, can accelerate the evolution of new forms of resistance - making them more important to the process than previously thought. (news-medical.net)
- Many of the most important resistance genes are found on plasmids, which are small, circular DNA molecules that live inside bacteria. (news-medical.net)
- Plasmids are capable of moving between bacteria and are usually thought of as being important 'vehicles' that transfer resistance genes between bacteria. (news-medical.net)
- This occurs because bacteria usually carry more than one copy of a plasmid, which allows resistance genes carried by plasmids to rapidly evolve new functions - in this case, the ability to degrade an antibiotic. (news-medical.net)
- Professor MacLean added: 'The conventional view of plasmids is that they act as important vehicles that transfer resistance genes between bacteria. (news-medical.net)
- Any of the conjugative or nonconjugative plasmids carrying genes responsible for antibiotic or antibacterial drug resistance among bacteria. (dictionary.com)
- For example, when tied to a gene promoter called araC, pGLO plasmids can be placed in bacteria, which subsequently glow green when in the presence of a sugar molecule called arabinose. (reference.com)
- During conjugation, gene transfer between bacteria is driven by a DNA plasmid, such as the F plasmid, in the bacterial cells. (reference.com)
- Natural conjugative plasmids expressed factors that induced planktonic bacteria to form or enter biofilm communities, which favour the infectious transfer of the plasmid. (nih.gov)
- For example, desirable genes can be inserted into plasmids in bacteria, which can then be used to produce large amounts of important proteins, such as insulin or antibiotics , or be used for waste treatment. (newworldencyclopedia.org)
- Plasmids that exist only as one or a few copies in each bacterium are, upon cell division, in danger of being lost in one of the segregating bacteria. (newworldencyclopedia.org)
- Some important groups of plasmids occurring in bacteria are given below. (newworldencyclopedia.org)
- In another words, an episome is a plasmid of bacteria or viral DNA that can integrate itself into the chromosomal DNA of the host organism (Figure 2). (newworldencyclopedia.org)
- There are two types of plasmid integration into a host bacteria: Non-integrating plasmids replicate as with the top instance, whereas episomes , the lower example, can integrate into the host chromosome . (wikipedia.org)
-  Plasmids can also provide bacteria with the ability to fix nitrogen . (wikipedia.org)
- In this study, we determined the distributions of the well-known NAPs Fis, H-NS, HU, IHF, and Lrp and the newly found NAPs MvaT and NdpA among the whole-sequenced 1382 plasmids found in Gram-negative bacteria. (hindawi.com)
- Join us on Saturday to learn how to extract and purify plasmids from bacteria. (meetup.com)
- Bacteria, plasmids, and phages. (worldcat.org)
- The isolation procedure is based on a modified alkaline lysis protocol and can be divided into the following steps:The bacteria are partially lysed, allowing the plasmid DNA to escape the cell wall into the supernatant. (sigmaaldrich.com)
- these plasmids can also be transfurred during sexual conjugation between bacteria. (studentdoctor.net)
- Cultures of bacteria containing low-copy-number plasmids amplified in the presence of chloramphenicol should be treated as if they contain high-copy-number plasmids when choosing the appropriate culture volumes for the QIAGEN-tip to be used. (qiagen.com)
- Furthermore the plasmid which you work with must be isolated from a dam+ bacteria strain. (jove.com)
- Plasmids usually occur naturally in bacteria , but are sometimes found in eukaryotic organisms (e.g., the 2-micrometre-ring in Saccharomyces cerevisiae ). (thefullwiki.org)
- Plasmids can be found in all three major domains , Archea , Bacteria and Eukarya . (thefullwiki.org)
- Plasmids also can provide bacteria with an ability to fix elemental nitrogen or to degrade calcitrant organic compounds which provide an advantage under conditions of nutrient deprivation. (thefullwiki.org)
- Next, the plasmids are inserted into bacteria by a process called transformation . (thefullwiki.org)
- Only bacteria which take up copies of the plasmid survive, since the plasmid makes them resistant. (thefullwiki.org)
- Now these bacteria can be grown in large amounts, harvested and lysed (often using the alkaline lysis method) to isolate the plasmid of interest. (thefullwiki.org)
- In this case, researchers grow bacteria containing a plasmid harboring the gene of interest. (thefullwiki.org)
- New thermosensitive plasmid for gram-positive bacteria. (asm.org)
- However, the genes of plasmids can confer advantages on bacteria living in stressful environments. (ukessays.com)
- Copy number depends on how actively transcribed the plasmids origin of replication is. (news-medical.net)
- If it is extremely active, plasmids can undergo several replication cycles during a single cell cycle. (news-medical.net)
- Other naturally occurring plasmids include those carrying the sex factor (F), which allows the cell to perform bacterial conjugation, replication origins and maintenance functions. (news-medical.net)
- Plasmids are a popular choice of vector as they are one of the most efficient ways to propagate DNA, they have an independent replication cycle and possess their own origin of replication. (news-medical.net)
- Plasmids Plasmids are extra-chromosomal, covalently closed circular (CCC) molecules of double stranded (ds) DNA that are capable of autonomous replication. (bookrags.com)
- The autonomous replication of the plasmid within the bacterial cells makes it possible to produce large numbers of copies of the recombinant DNA molecule for experimental manipulation or commercial purposes (such as the production of large amounts of insulin). (britannica.com)
- My plasmids have only the 2u replication origin though, also pretty common. (bio.net)
- Although the enzymes involved in DNA replication of plasmids are the same as those used for the chromosome, some plasmids are copied in a different way than the chromosome. (dummies.com)
- For example, it may be possible to develop new drugs that will block plasmid replication. (news-medical.net)
- A plasmid is an extrachromosomal, extranuclear DNA molecule occurring free in the cytoplasm and capable of autonomous replication (independently of the chromosomal DNA). (newworldencyclopedia.org)
- But the initiation of replication and the distribution of plasmid copies to the daughter cells is controlled by plasmid genes . (newworldencyclopedia.org)
- Thus, every plasmid contains at least one DNA sequence that serves as an origin of replication, or ori (a starting point for DNA replication), which enables the plasmid DNA to be duplicated independently from the chromosomal DNA (Figure 4) and to be distributed to the daughter cells. (newworldencyclopedia.org)
- Artificial plasmids are widely used as vectors in molecular cloning , serving to drive the replication of recombinant DNA sequences within host organisms. (wikipedia.org)
-  The term's early usage included any bacterial genetic material that exists extrachromosomally for at least part of its replication cycle, but because that description includes bacterial viruses, the notion of plasmid was refined over time to comprise genetic elements that reproduce autonomously. (wikipedia.org)
- In order for plasmids to replicate independently within a cell, they must possess a stretch of DNA that can act as an origin of replication . (wikipedia.org)
- A typical bacterial replicon may consist of a number of elements, such as the gene for plasmid-specific replication initiation protein (Rep), repeating units called iterons , DnaA boxes, and an adjacent AT-rich region. (wikipedia.org)
-  Smaller plasmids make use of the host replicative enzymes to make copies of themselves, while larger plasmids may carry genes specific for the replication of those plasmids. (wikipedia.org)
- 3. A composition according to claim 1 wherein said plasmid comprises a polydeoxyribonucleotide containing more than one origin of replication that is recognized by human DNA polymerase. (google.com)
- 6. A composition according to claim 3 wherein the presence in said plasmid of multiple copies of an origin of replication facilitates efficient replication of said plasmid in human cells. (google.com)
- However, a number of plasmids do not contain the gene encoding relaxase ( mobA ) and it is necessary to use the analysis of the replication genes in this case. (frontiersin.org)
- These plasmids carried conserved IncHI5 backbones composed of repHI5B and a repFIB -like gene (replication), parABC (partition), and tra1 (conjugal transfer). (frontiersin.org)
- When low-copy-number plasmids containing the pMB1 or ColE1 origin of replication are prepared using QIAGEN Plasmid Purification Kits , plasmid DNA yields can be improved by adding chloramphenicol to the culture medium (170 mg/liter) to amplify copy numbers. (qiagen.com)
- Please see FAQ 350 for origins of replication and copy numbers of various plasmids. (qiagen.com)
- Same Replication plasmids are incompatible. (protocol-online.org)
- Replication-defective mutants of plasmid ColE1 were isolated from a chimeric plasmid formed by ligating a temperature-sensitive replication derivative of pSC101, pHSG1, with a ColE1-Tn3-containing plasmid. (unboundmedicine.com)
- Hashimoto-Gotoh T, Inselburg J. Isolation and characterization of replication-deficient mutants of ColE1 plasmids. (unboundmedicine.com)
- TY - JOUR T1 - Isolation and characterization of replication-deficient mutants of ColE1 plasmids. (unboundmedicine.com)
- The sequences of the genes involved in the replication of pCD1 are highly homologous to those of the cognate plasmids in Y. pseudotuberculosis and Y. enterocolitica , but their localization within the plasmid differs markedly from those of the latter. (asm.org)
- Plasmids are considered transferable genetic elements, or " replicons ", capable of autonomous replication within a suitable host. (thefullwiki.org)
- Here we describe the current knowledge and understanding of the structure of iteron-containing replicons, the structure of the iteron plasmid encoded replication initiation proteins, and the molecular mechanisms for iteron plasmid DNA replication initiation. (asmscience.org)
- Nucleotide sequence of the region of the origin of replication of the broad host range plasmid RK2. (asmscience.org)
- Nine unique repeating sequences in a region essential for replication and incompatibility of the mini-F plasmid. (asmscience.org)
- 1984. P1 plasmid replication: replicon structure. (asmscience.org)
- DNA and protein interactions in the regulation of plasmid replication. (asmscience.org)
- Replication functions encoded by the plasmid pSC101. (asmscience.org)
- Replication initiator protein RepE of mini-F plasmid: functional differentiation between monomers (initiator) and dimers (autogenous repressor). (asmscience.org)
- The plasmid RK2 initiation protein binds to the origin of replication as a monomer. (asmscience.org)
- Modified plasmids also carry a marker gene. (news-medical.net)
- A standard cloning procedure begins with the ligation of a plasmid vector, with a marker gene, and the target DNA fragment. (news-medical.net)
- Plasmids are extremely valuable tools in the fields of molecular biology and genetics, specifically in the area of genetic engineering ( q.v. ). They play a critical role in such procedures as gene cloning, recombinant protein production ( e.g., of human insulin), and gene therapy research. (britannica.com)
- A foreign DNA element (such as the gene for insulin) is then spliced into the plasmid. (britannica.com)
- In DNA cloning, a DNA fragment that contains a gene of interest is inserted into a cloning vector or plasmid . (accessexcellence.org)
- The plasmid carrying genes for antibiotic resistance, and a DNA strand, which contains the gene of interest, are both cut with the same restriction endonuclease . (accessexcellence.org)
- The plasmid is opened up and the gene is freed from its parent DNA strand. (accessexcellence.org)
- The opened plasmid and the freed gene are mixed with DNA ligase, which reforms the two pieces as recombinant DNA. (accessexcellence.org)
- I think Rose & Broach (Gene Expression Technology pp 234-279, ed Goeddel 1990) is a good starting point for the 2u plasmid stability literature. (bio.net)
- I just found the pEU720 (described in Froehlich and Scott, Gene 108 (1991), 99-101) and the F-plasmid. (bio.net)
- The mitochondrial gene oxi1, carried on a bacterial plasmid, has been used to transform the mitochondria of a yeast strain lacking mtDNA (rho0). (pnas.org)
- Plasmid welcomes topics such as horizontal gene transfer, including antibiotic resistance transfer, and molecular aspects of microbial ecology. (elsevier.com)
- The pGLO plasmid is a small circular piece of DNA that contains the gene to produce green fluorescent protein in the model organism. (reference.com)
- Within the pGLO plasmid, the gene that codes for GFP is typically linked to a gene promoter of interest. (reference.com)
- This plasmid carries the E. coli malE gene, encoding the maltose binding protein (MBP)(3), fused in-frame to the coding region of the Sce VMA intein-chitin binding domain (55 kDa)(1,4). (neb.com)
- This term is no longer commonly used for plasmids, since it is now clear that a transposon (jumping gene or mobile genetic unit) makes a plasmid into an episome. (newworldencyclopedia.org)
-  This host-to-host transfer of genetic material is one mechanism of horizontal gene transfer , and plasmids are considered part of the mobilome . (wikipedia.org)
- Plasmids almost always carry at least one gene. (wikipedia.org)
- We found that larger plasmids frequently have NAP gene homologs. (hindawi.com)
- Plasmids with H-NS gene homologs had less G+C content. (hindawi.com)
- It should be noted that plasmids with the NAP gene homolog also carried the relaxase gene involved in the conjugative transfer of plasmids more frequently than did those without the NAP gene homolog, implying that plasmid-encoded NAP homologs positively contribute to transmissible plasmids. (hindawi.com)
- DNA sequence polymorphism was determined for the microcin V gene cluster encoded on the microcin V plasmids of 12 natural isolates of Escherichia coli. (umass.edu)
- With respect to therapeutic compositions for gene therapy, the DNA is provided typically in the form of a plasmid for complexing with the cationic amphiphile. (google.com)
- Novel and highly effective plasmid constructs are also disclosed, including those that are particularly effective at providing gene therapy for clinical conditions complicated by inflammation. (google.com)
- Plasmid DNA is a key component in the production of most gene and gene-enabled cell therapies. (biospace.com)
- The PathDetect Control Reporter Plasmid allows you to determine the direct or indirect involvement of new gene products, growth factors, and drug candidates in specific pathways. (agilent.com)
- The plasmid is designed to reduce the cost of AAV manufacturing and subsequent gene therapy applications, while helping Aldevron's clients meet regulatory requirements. (biospace.com)
- Thousands of clients use Aldevron-produced plasmids, RNA and gene editing enzymes for projects ranging from discovery research to clinical trials to commercial applications. (biospace.com)
- However, the design and construction of plasmid donors for each gene can be laborious and prone to troubleshooting. (genetics.org)
- However, ssDNA donors are limited to small insertions, and, like plasmid donors, must be designed and generated for each gene that is targeted by HDR. (genetics.org)
- This is now possible utilizing bionic electrode arrays for focal delivery of naked (plasmid) DNA via gene electrotransfer. (nature.com)
- An array with eight 300 μm platinum ring electrodes configured as a cochlear implant bionic interface was used to transduce HEK293 cell monolayers with a plasmid-DNA green fluorescent protein (GFP) reporter gene construct. (nature.com)
- The complete nucleotide sequence and gene organization of the three virulence plasmids from Yersinia pestis KIM5 were determined. (asm.org)
- With this method the cloned target gene can be altered by substitution, deletion or insertion of a few bases directly into a plasmid. (jove.com)
- The site directed mutagenesis of whole plasmids explained in this video is a mutagenesis method which allows you to alter a cloned target gene by substitution, deletion or insertion of a few bases directly into a plasmid. (jove.com)
- Plasmids can be considered to be part of the mobilome , since they are often associated with conjugation , a mechanism of horizontal gene transfer . (thefullwiki.org)
- Rather, plasmids provide a mechanism for horizontal gene transfer within a population of microbes and typically provide a selective advantage under a given environmental state. (thefullwiki.org)
- The gene to be replicated is inserted into copies of a plasmid containing genes that make cells resistant to particular antibiotics and a multiple cloning site (MCS, or polylinker), which is a short region containing several commonly used restriction sites allowing the easy insertion of DNA fragments at this location. (thefullwiki.org)
- The GeneArt™ gene synthesis platform can design plasmid vectors from modular, easily exchangeable fragments encoding regulatory and functional elements or from custom-made elements. (thermofisher.com)
- For example, strains such as Escherichia coli can have three different small plasmids in many copies, and one large single copy plasmid. (news-medical.net)
- Plasmids which replicate in E. coli are one of the most common choices of vector in DNA cloning. (news-medical.net)
- Members of Enterobacteriaceae family, for example, Escherichia coli or Klebsiella pneumoniae pose the biggest threat regarding plasmid-mediated resistance in hospital- and community-acquired infections. (wikipedia.org)
- In E. coli Plasmid Vectors, experienced bench researchers describe their proven techniques for the manipulation of recombinant plasmids utilizing this popular bacterial host. (springer.com)
- The authors describe readily reproducible methods for cloning DNA into plasmid vectors, transforming plasmids into E. coli, and analyzing recombinant clones. (springer.com)
- Comprehensive and highly practical, E. coli Plasmid Vectors offers those new to the field a basic guide to the use of plasmid vectors in the cloning host E. coli, and those more experienced researchers a broad-ranging, proven array of successful techniques. (springer.com)
- Dear all, does anyone of you know of a plasmid (for E coli or Bacillus) that exists only as one copy in the cell? (bio.net)
- Studies on transformation of Escherichia coli with plasmids. (nih.gov)
- Factors that affect the probability of genetic transformation of Escherichia coli by plasmids have been evaluated. (nih.gov)
- aureus and Escherichia coli harbouring reference plasmids pUC18 and RP4, and resistant R plasmids from clinical isolates of Enterococcus faecalis, Bacillus cereus, E. (thefreedictionary.com)
- Can phage defence maintain colicin plasmids in Escherichia coli? (umass.edu)
- We examined the role of plasmid-based phage defence in maintaining plasmids, using colicin plasmids in Escherichia coli as a model system. (umass.edu)
- 1995b Can phage defence maintain colicin plasmids in Escherichia Coli? (umass.edu)
- GenElute™ Plasmid Miniprep Kit offers a simple, rapid, and cost-effective method for isolating up to 20 µg plasmid DNA from E. coli cultures. (sigmaaldrich.com)
- The Thermo Scientific MagJET Plasmid DNA Kit is designed for fast and efficient purification of plasmid DNA from overnight cultures of E. coli cultures. (fishersci.com)
- A plasmid was introduced into competent Escherichia coli (E.coli) cells by electroporation, plated out on a solid medium and left to divide and multiply. (ukessays.com)
- In this experiment, the plasmid pUC19 was introduced into competent Escherichia coli (E.coli) cells. (ukessays.com)
- This, thereby, makes it easier for the plasmid to enter the E.coli cells. (ukessays.com)
- The negative control is made using E.coli without the plasmid while the positive control contains E.coli with the plasmid. (ukessays.com)
- but linear plasmids have also been reported. (newworldencyclopedia.org)
- A book that constitutes the first attempt to comprehensively assemble current knowledge of different types of such elements, highlight recent developments in the field, and challenge the distinction between viruses and linear plasmids. (foyles.co.uk)
- Linear plasmids of microbes represent a heterogenous group of extrachromosomal genetic elements initially assumed to be rare and peculiar. (foyles.co.uk)
- Eighty progeny were analyzed from a cross in which the maternal (stromal) parent possessed three linear plasmids, designated Callan-a (7.5 kb), Aubonne-a (2.1 kb) and Bergell (2.0 kb), and the paternal parent had one plasmid, Aubonne-b (2.1 kb). (genetics.org)
- Most of the resistance plasmids are conjugative, meaning that they encode all the needed components for the transfer of the plasmid to other bacterium. (wikipedia.org)
- Schematic drawing of a bacterium with plasmids enclosed. (newworldencyclopedia.org)
- Illustration of a bacterium showing chromosomal DNA and plasmids. (wikipedia.org)
-  Plasmids are transmitted from one bacterium to another (even of another species) mostly through conjugation . (wikipedia.org)
- Plasmids can jump from one bacterium to another, spreading that resistance. (medicalxpress.com)
- Another complication is that certain R plasmids can carry up to 10 resistance genes to different antibiotics, and these plasmids often contain genes which allow bacterial conjugation to occur. (news-medical.net)
- This general connection between conjugation and biofilms suggests that medically relevant plasmid-bearing strains are more likely to form a biofilm. (nih.gov)
- Episomes, Plasmids, Insertion Sequences and Transposons Episomes, plasmids, insertion sequences and transposons are deoxyribonucleic acid (DNA) which can exist independent of the main, or genomic, DNA. (bookrags.com)
- Plasmids are circular deoxyribonucleic acid (DNA) molecules that replicate independently of the bacterial chromosome . (britannica.com)
- The extent of a deletion was determined by analyzing restriction endonuclease-generated deoxyribonucleic acid fragments of the ColE1 plasmid component of the chimeras by both agarose and polyacrylamide gel electrophoresis. (unboundmedicine.com)
- Coloured atomic force micrograph of numerous pGL3 plasmids of DNA (deoxyribonucleic acid). (sciencephoto.com)
- These experiments led to the genetic definition of two conjugative plasmids-SCP1 and SCP2-that were deduced to be present in an autonomous state in the wild-type A3(2) strain and to be lost, or in the case of SCP1 sometimes chromosomally integrated, in various of its derivatives. (springer.com)
- Bibb, M. J., and Hopwood, D. A., 1981, Genetic studies of the fertility plasmid SCP2 and its SCP2' variants in Streptomyces coelicolor A3(2), J. Gen. Microbiol . (springer.com)
- Plasmid , in microbiology, an extrachromosomal genetic element that occurs in many bacterial strains. (britannica.com)
- Plasmids are well suited to genetic engineering in other ways. (britannica.com)
- oxi1 genetic information in these "synthetic rho-" strains could be expressed in diploids either after "marker rescue" by recombination with rho+ mtDNA carrying an appropriate oxi1 point mutation or in trans during the growth of diploids heteroplasmic for both the plasmid-derived oxi1 sequences and rho+ mtDNA with oxi1 deleted. (pnas.org)
- Plasmid publishes original research on genetic elements in all kingdoms of life with emphasis on maintenance, transmission and evolution of extrachromosomal elements . (elsevier.com)
- While chromosomes are large and contain all the essential genetic information for living under normal conditions, plasmids are usually very small and contain only additional genes that may be useful in certain situations or conditions. (wikipedia.org)
-  Later in 1968, it was decided that the term plasmid should be adopted as the term for extrachromosomal genetic element,  and to distinguish it from viruses, the definition was narrowed to genetic elements that exist exclusively or predominantly outside of the chromosome and can replicate autonomously. (wikipedia.org)
- Circularized plasmid DNA remains extra-chromosomal and can drive sustained episomal expression, while linearized DNA may be integrated into the chromosomal DNA, enabling targeted genetic modifications. (nature.com)
- Iteron-containing plasmids are model systems for studying the metabolism of extrachromosomal genetic elements in bacterial cells. (asmscience.org)
- Natural plasmids must be genetically modified before being used as a vector for cloning. (news-medical.net)
- This makes the plasmid a more flexible cloning vector. (news-medical.net)
- this means that surviving colonies have been transformed by plasmid vector, allowing them to grow in that environment. (news-medical.net)
- David Wessner reviews Vector NTI Suite, an INternet integrated software tool that can catalog, analyze, and design complex plasmids for molecular biologists. (sciencemag.org)
- Activity 2 will help students conceptualize the mechanics involved in cutting and ligating DNAs into a plasmid vector with "sticky ends" of complementary DNA base pairs. (accessexcellence.org)
- The vector DNA (plasmid) containing the "new" genes must now be inserted into host cells. (accessexcellence.org)
- If the plasmid vector you need is not available, either commercially or through your research network, we can help. (thermofisher.com)
- Then the genes have to be inserted into a vector such as a plasmid. (ukessays.com)
- Plasmids responsible for "fertility" (or "chromosome mobilizing ability" [Cma]) (24) were also identified genetically in some other strains, including Streptomyces rimosus (18), Streptomyces lividans (29), Streptomyces erythreus (now called Saccharopolyspora erythrea ) (15), Streptomyces venezuelae (17), and Streptomyces ambofaciens (66). (springer.com)
- In addition, since the plasmids that carry ESBL genes also commonly encode resistance determinants for many other antibiotics, ESBL strains are often resistant to many non-beta-lactam antibiotics as well, leaving very few options for the treatment. (wikipedia.org)
- Like the mtDNA sequences retained in natural rho- strains, the plasmid DNA in the transformants was reiterated into concatemers whose size was indistinguishable from that of wild-type mtDNA. (pnas.org)
- These plasmids are interesting because of their size, they range between 100 and 1800 Kb, and in some strains they represent about 50% of the bacterial genome. (elsevier.com)
- Here, we characterized 44 plasmids isolated from five permafrost Acinetobacter lwoffii strains, examined their relationship with plasmids of modern Acinetobacter strains and identified groups of related plasmids. (frontiersin.org)
- The presence of plasmids in clinical strains that are closely related to those of environmental permafrost strains provides evidence of the origin of the former from the latter. (frontiersin.org)
- Our system uses gRNA-based identification markers that allow messages to be addressed to specific strains via Cas9-mediated cleavage of messages sent to the wrong recipient, which we show reduces plasmid transfer by four orders of magnitude. (caltech.edu)
- They are designed so target restriction fragments, cut by specific restriction enzymes, have a unique location in that plasmid. (news-medical.net)
- Many modified plasmids have a variety of unique restriction fragment sites, formed by unique combinations of restriction enzymes. (news-medical.net)
- I used sucrose gradients to isolate my plasmid preps - got good enough DNA to produce good restriction patterns, clone specific fragments etc. (bio.net)
- In such procedures, a plasmid is cut at a specific site (or sites) using enzymes called restriction endonucleases. (britannica.com)
- The oxi1 sequences in the transformants were surrounded by restriction sites derived from the plasmid that were not present in wild-type mtDNA. (pnas.org)
- Plasmid DNA is ready for immediate use in restriction enzyme digestion (Figure 1) cloning, PCR, transcription, conventional and automated sequencing (Figure 2) . (sigmaaldrich.com)
- Plasmid DNA purified with GenElute Plasmid Miniprep Kit is suitable for restriction enzyme digestions. (sigmaaldrich.com)
- Restriction digestions of pUC19 purified from JM109 using the GenElute Plasmid Miniprep Kit. (sigmaaldrich.com)
- a 2-kilobase restriction fragment that contains RLF3 was subcloned into plasmid pBR322 prior to further characterization. (reverso.net)
- un fragment de restriction de 2 kilobase, contenant le RLF3, a été subcloné en un plasmide pBR322 avant de procéder à sa caractérisation. (reverso.net)
- Thereafter you can use this plasmid as template for a series of mutations in which this restriction site will be deleted by insertion of the new mutagenic primers. (jove.com)
- Once your plasmid backbone has been amplified and purified you must digest it with the flanking restriction enzymes that you used to cut your two parts that you are assembling. (igem.org)
- It is very common for the resistance genes or entire resistance cassettes to be re-arranged on the same plasmid or be moved to a different plasmid or chromosome by means of recombination systems. (wikipedia.org)
- A plasmid that is attached to the cell membrane or integrated into the bacterial chromosome is called an episome ( q.v. ). (britannica.com)
- Plasmids contain only nonessential genes and replicate independent of the chromosome. (dummies.com)
- A few types of plasmids can also insert into the host chromosome, and these integrative plasmids are sometimes referred to as episomes in prokaryotes. (wikipedia.org)
- The definition of a resistance plasmid is a small element outside the chromosome that carries DNA information that fights against antibiotic drugs. (yourdictionary.com)
- A plasmid is a small, circular, self-replicating DNA molecule which differs from the bacterial chromosome. (ukessays.com)
Antibiotic resistance genes6
- For example, plasmids can contain antibiotic resistance genes, posing a risk to public health. (news-medical.net)
- Plasmid-mediated resistance is the transfer of antibiotic resistance genes which are carried on plasmids. (wikipedia.org)
- Plasmids often carry multiple antibiotic resistance genes, contributing to the spread of multidrug-resistance (MDR). (wikipedia.org)
- Resistance plasmids by definition carry one or more antibiotic resistance genes. (wikipedia.org)
- The antibiotic resistance genes found on the plasmids confer resistance to most of the antibiotic classes used nowadays, for example, beta-lactams, fluoroquinolones and aminoglycosides. (wikipedia.org)
- Plasmids are pieces of independent DNA that often carry multiple antibiotic resistance genes. (medicalxpress.com)
- http://www.protocol-online.org/prot/Molecular_Biology/Plasmid/Miniprep/ A miniprep procedure using diatomaceous earth to bind DNA during purification and washing. (wikipedia.org)
- The term plasmid was first introduced by the American molecular biologist Joshua Lederberg in 1952 to describe any extrachromosomal hereditary determinant. (newworldencyclopedia.org)
- This work has contributed to a better understanding of the role of plasmids in the rhizobial life cycle, through the study of plasmid encoded functions, and the analysis of the molecular mechanisms that allow conjugative transfer of these plasmids. (elsevier.com)
- The presence of these non-antimicrobial-drug resistance R plasmid DNA sequences in SGI1 constitutes a molecular signature that firmly establishes the aquaculture origin of the florfenicol resistance and the tetR/ tetG genes in the S. (thefreedictionary.com)
- Circular plasmids are commonly used as donor DNA for HDR because they can carry a large DNA insert (≤ 10 kb) and homology arms corresponding to the target locus are added by molecular cloning. (genetics.org)
- The RapidPURE™ Plasmid Mini Kit is designed to isolate up to 20 μg of plasmid DNA from 1.5 mL of an overnight bacterial culture for use in routine molecular biology applications such as fluorescent and radioactive sequencing and cloning. (mpbio.com)
- The term plasmid was first introduced by the American molecular biologist Joshua Lederberg in 1952. (thefullwiki.org)
- Larger Plasmids are a bit difficult to mutate with this method and may take some patience and adjustment of the thermocycling conditions and/or competent cells. (jove.com)
- A plasmid is an extra chromosomal DNA molecule separate from the chromosomal DNA which is capable of replicating independently from the chromosomal DNA. (thefullwiki.org)
- Kits are available from varying manufacturers to purify plasmid DNA, which are named by size of bacterial culture and corresponding plasmid yield. (wikipedia.org)
- Purify all sizes and types of plasmid, even BAC DNA, since the crude lysate can be filtered to avoid plasmid shearing. (sigmaaldrich.com)
- The CosMCPrep system is a versatile plasmid purification procedure designed to purify a variety of high- and low-copy number template types. (beckman.com)
- Learn how to purify transfection grade plasmid DNA with the Invitrogen PureLink HiPure Plasmid Filter Maxiprep Kit. (thermofisher.com)
- Plasmid pPCP1 (9,610 bp) has a GC content of 45.3% and encodes two previously known virulence factors, an associated protein, and a single copy of IS 100 . (asm.org)
- The present invention relates to Mycobacterium ulceran virulence plasmid , pMUM001 and particularly to a cluster of genes carried by this plasmid that encode polyketide synthases (PKSs) and polyketide-modifying enzymes necessary and sufficient for mycolactone biosynthesis. (reverso.net)
- La présente invention se rapporte à un plasmide de virulence de Mycobacterium ulcerans, pMUM001, et notamment à une famille de gènes portés par ce plasmid et codant les polycétide-synthases (PKS) et les enzymes modificatrices de polycétides nécessaires et suffisantes pour la synthèse de la mycolactone. (reverso.net)
Presence of multiple plasmids2
- Competition with both transforming and non-transforming plasmids indicates that each cell is capable of taking up many DNA molecules, and that the establishment of a transformation event is neither helped nor hindered significantly by the presence of multiple plasmids. (nih.gov)
- An important feature of Acinetobacter is also the presence of multiple plasmids in the same strain. (frontiersin.org)
- A set of conditions is described under which about one in every 400 plasmid molecules produces a transformed cell. (nih.gov)
- A new study led by scientists at the University of Oxford has found that small DNA molecules known as plasmids are one of the key culprits in spreading the major global health threat of antibiotic resistance. (news-medical.net)
- Currently, however, the term plasmid is restricted only to those accessory DNA molecules that are found in addition to the main chromosomes. (newworldencyclopedia.org)
- Functional vRNP molecules could be generated either by infection with helper virus or by cotransfection of protein expression plasmids encoding PB1, PB2, PA, or nucleoproteins ( 11 , 13 - 15 ). (pnas.org)
- After the addition of acetate-containing neutralization buffer the large and less supercoiled chromosomal DNA and proteins precipitate, but the small bacterial DNA plasmids stay in solution. (wikipedia.org)
- Plasmids were first clearly implicated in this conjugative process in the most studied strain, Streptomyces coelicolor A3(2), when certain derivatives of the wild-type isolate were found to differ in their "fertility" properties-that is, in the frequency with which they generated chromosomal recombinants when mated with various other derivatives-and this ability was inherited "infectiously" (2, 28, 72). (springer.com)
- A plasmid is a small, extrachromosomal DNA molecule within a cell that is physically separated from chromosomal DNA and can replicate independently. (wikipedia.org)
- A plasmid is a loop of DNA that can exist in a cell independently of the cell's chromosomal DNA. (sciencephoto.com)
- Plasmids are important for bacterial evolution and adaptation to the changing environment, as they carry genes which carry beneficial traits for the bacterial cell. (news-medical.net)
- In nature, plasmids often carry genes that benefit the survival of the organism and confer selective advantage such as antibiotic resistance . (wikipedia.org)
- Recent studies have shown that some plasmids carry genes encoding NAP homologs, which play important roles in transcriptional regulation networks between plasmids and host chromosomes. (hindawi.com)
- Plasmids may carry genes that provide resistance to naturally occurring antibiotics in a competitive environmental niche, or alternatively the proteins produced may act as toxins under similar circumstances. (thefullwiki.org)
- They conducted whole-genome analyses on the samples to study the bacterial plasmids, or rings of DNA, that can confer resistance to antibiotics. (eurekalert.org)
- The majority of samples they studied from the pipes and sewers tested positive for bacterial plasmids that confer resistance to carbapenems, the researchers report this week in mBio . (eurekalert.org)
- The new findings add to a growing body of evidence suggesting that the conduits of hospital wastewater serve as a vast and resilient reservoir for plasmids that can confer the genes responsible for antibiotic resistance. (eurekalert.org)
- Experimental data indicated that the possession of a colicin plasmid can confer limited protection against bacteriophages. (umass.edu)
- PCR based analysis of genomic DNA from muscle samples, taken from at or around the injection site 436 days post vaccination, led the company to conclude that integration of plasmid DNA into the fish genome is extremely unlikely. (europa.eu)
- In this method, simultaneous cutting of a circular donor plasmid and a genomic target site by a nuclease results in integration of linearized insert DNA into the genomic cut site by nonhomologous end joining (NHEJ). (genetics.org)
- We analyzed the differences in information capacity between prokaryotic chromosomes, genomic islands (GI), phages, and plasmids. (wur.nl)
Alkaline lysis method1
- The Genopure Plasmid Midi Kit prepares highly purified plasmid DNA in medium quantities using a modified alkaline lysis method. (sigmaaldrich.com)
- We have developed an eight-plasmid DNA transfection system for the rescue of infectious influenza A virus from cloned cDNA. (pnas.org)
- In this plasmid-based expression system, viral cDNA is inserted between the RNA polymerase I (pol I) promoter and terminator sequences. (pnas.org)
- The novel receptor is encoded for by cDNA carried on plasmid pGEM-hRARn, which has been desposited with the American Type Culture Collection for patent purposes. (reverso.net)
Nonprofit plasmid repository2
- Thus, one can conclude that there is not and cannot exist a universal system based on the analysis of only one plasmid protein, rep, mob , or another. (frontiersin.org)
- Using this method in cultured S2R+ cells, we efficiently tagged four endogenous proteins with the bright fluorescent protein mNeonGreen, thereby demonstrating that an existing collection of CRISPaint universal donor plasmids is compatible with insect cells. (genetics.org)
- In addition to 5 previously described genes, such as murine toxin, capsule antigen, capsule anchoring protein, etc., 30 homologues to genes of several bacterial species were found in this plasmid, and another 44 open reading frames without homology to any known or hypothetical protein in the databases were predicted. (asm.org)
- Our original approach was to use plasmid containing the P1010 insert, coding for the ccdB protein generator. (igem.org)
- high copy number plasmids can be more than 100 copies per cell, whereas others are limited to very few copies per cell. (news-medical.net)
- Some cells have plasmids, which are smaller circular DNA , in addition to chromosomes. (studentdoctor.net)
- Plasmids are important as they can replicate and express genes despite being outside the organism's chromosomes. (sciencephoto.com)
- There was an association between relative entropy and AT content in chromosomes, phages, plasmids and GIs with the strongest association being in phages. (wur.nl)
- Conclusions: We argue that relative entropy differences reflect how plasmids, phages and GIs interact with microbial host chromosomes and that all these biological entities are, or have been, subjected to different selective pressures. (wur.nl)
- A bacterial plasmid is a small DNA molecule that are commonly used bacterial cloning. (news-medical.net)
- Such single-copy plasmids have systems that are oriented toward actively distributing a copy to both daughter cells. (newworldencyclopedia.org)
- Comparison of non-integrating plasmids (top) and episomes (bottom) . (newworldencyclopedia.org)
- Transformation efficiency declines linearly with increasing plasmid size. (nih.gov)
- pMYB5 can be used to test plasmid transformation, cell culture, induction and purification procedures. (neb.com)
- In the laboratory, plasmids may be introduced into a cell via transformation . (wikipedia.org)
- AFAIK would be better to redesign the plasmid, unless the GFP is on the same plasmid, it would be useless for checking transformation efficiency if it has much higher transformation efficiency than your target. (protocol-online.org)
- [ 1 ] Microbial transformation with plasmid DNA is neither parasitic nor symbiotic in nature, since each implies the presence of an independent species living in a commensal or detrimental state with the host organism. (thefullwiki.org)
- Transformation can also be carried out by introducing a plasmid into a bacterial cell. (ukessays.com)
- Would it be possible to co-transfect my HEK293 with another plasmid that has a GFP insert but with another antibiotic resistance? (protocol-online.org)
Size of plasmids2
Wherein said plasmid3
- c) removing said detergent from said solution of step (b) to provide a solution of serum-stable plasmid-lipid particles, wherein said plasmid is encapsulated in a lipid bilayer and said particles are resistant to degradation in serum, and wherein the particles have a diameter ranging from about 50 to about 150 nm. (google.com)
- 11. A method in accordance with claim 10, wherein said plasmid-lipid particle comprises a plasmid, DODAC, POPC and a PEG-Ceramide selected from the group consisting of PEG-Cer-C 20 and PEG-Cer-C 14 . (google.com)
- 4. A composition according to claim 3 wherein said plasmid further facilitates interaction of said polymerase with itself. (google.com)
- The cloning plasmids contain a site for a DNA insert. (news-medical.net)
- These sites flank the cloning site in the plasmid. (news-medical.net)
- Process by which a plasmid is used to import recombinant DNA into a host cell for cloning. (accessexcellence.org)
- Ultrapure DNA purified with HiSpeed Plasmid Kits yields excellent results in all applications, from cloning and sequencing to transfection. (bio-medicine.org)
- For details and precautions on the use of chloramphenicol when culturing plasmids, please refer to standard manuals for cloning procedures (e.g. (qiagen.com)
- Using primers which match the biobrick cloning sites in the reverse orientation, we produce PCR product containing the plasmid backbone with no insert and no uncut, circular plasmid. (igem.org)
- Since the quality of this construction plasmid is a key determiner in the success of three antibiotic cloning, substantial effort has been put into optimizing the reactions. (igem.org)
- Derivatives of the thermosensitive plasmid convenient for cloning purposes have been constructed. (asm.org)
- Plasmids can only exist and replicate within a cell, where it uses host cell machinery. (news-medical.net)
- Plasmid-lipid particles which are useful for transfection of cells in vitro or in vivo are described. (google.com)
- The Genopure Plasmid Midi Kit prepares transfection-grade plasmid DNA in medium quantities (up to 100 μg plasmid) from bacterial cultures. (sigmaaldrich.com)
- Seventy-two hours after the transfection of eight expression plasmids into cocultured 293T and MDCK cells, the virus yield in the supernatant of the transfected cells was between 2 × 10 5 and 2 × 10 7 infectious viruses per milliliter. (pnas.org)
- My current plasmid of interest was constructed in another lab and doesn't contain the GFP tag, so I wouldn't be able to see transfection efficiency post-transfection. (protocol-online.org)
- What would the transfection efficiency be if I had two plasmids? (protocol-online.org)
- You can't assume that the transfection efficiencies will be the same for both plasmids - you can do immuno to assess the rates for each. (protocol-online.org)
- The bigger the plasmid, the poorer the transfection usually. (protocol-online.org)
- You would need the same selection marker on both plasmids to make co-transfection more likely. (protocol-online.org)
- prabhubct: If you only select for one plasmid the other will be treated by the cells in a simiar manner to a transient transfection, and will be lost after a few days. (protocol-online.org)
Agarose gel analysis1
- 1 μg of plasmid was completely digested with 1 U Msp I for 2 hours at +37°C, as shown by agarose gel analysis. (sigmaaldrich.com)
- I would go for electroporation for large plasmids. (protocol-online.org)
- Based on the analysis, 19 major groups (lineages) of plasmids were identified, of which more than half were small plasmids. (frontiersin.org)
- pMYB5 is a control plasmid for the IMPACT™ Kit (NEB #E6901) (1,2). (neb.com)
- Construction plasmid is uncut backbone plasmid DNA from the three different plasmid backbones used for three antibiotic assembly (pSB1C3, pSB1K3, and pSB1T3). (igem.org)
- The size of the plasmid varies from 1 to over 200 k bp ,  and the number of identical plasmids in a single cell can range anywhere from one to thousands under some circumstances. (wikipedia.org)
- This requires a 'ligase' enzyme and forms a recombinant plasmid. (news-medical.net)
- Plasmids + copies of the DNA fragment produce quantities of recombinant DNA. (accessexcellence.org)
- Additionally, plasmids automatically amplify the number of copies of these new and improved resistance genes. (news-medical.net)
- Depending on the size of the plasmids, the number of copies of the same plasmid in a single cell varies from one to several hundreds. (newworldencyclopedia.org)
- Plasmids serve as important tools in genetics and biotechnology labs, where they are commonly used to multiply (make many copies of) or express particular genes. (thefullwiki.org)