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 ...
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
The Relaxosome is the complex of proteins that facilitates plasmid transfer during bacterial conjugation. The proteins are encoded by the tra operon on a fertility plasmid in the region near the origin of transfer, oriT. The most important of these proteins is relaxase, which is responsible for beginning the conjugation process by cutting at the nic site via transesterification. This nicking results in a DNA-Protein complex with the relaxosome bound to a single strand of the plasmid DNA and an exposed 3 hydroxyl group. Relaxase also unwinds the plasmid being conjugated with its helicase properties. The relaxosome interacts with integration host factors within the oriT. Other genes that code for relaxosome components include TraH, which stabilizes the relaxosomes structural formation, TraI, which encodes for the relaxase protein, TraJ, which recruits the complex to the oriT site, TraK, which increases the nicked state of the target plasmid, and TraY, which imparts single-stranded DNA ...
TY - JOUR. T1 - The first high frequency of recombination-like conjugal transfer from an integrated origin of transfer sequence in Bacillus subtilis 168. AU - Itaya, Mitsuhiro. AU - Hasegawa, Miki. AU - Tomita, Masaru. AU - Sato, Mitsuru. PY - 2018/1/1. Y1 - 2018/1/1. N2 - Bacillus subtilis 168 was developed as a genome vector to manipulate large DNA fragments. The system is based on the inherent natural transformation (TF) activity. However, DNA size transferred by TF is limited up to approximately 100 kb. A conjugal transfer system capable of transferring DNA fragments considerably larger than those transferred by TF was developed. A well-defined oriT110 sequence and a cognate relaxase gene from the pUB110 plasmid were inserted into the xkdE gene of the B. subtilis genome. Transfer of antibiotic resistance markers distant from the oriT110 locus to the recipient B. subtilis occurred only in the presence of pLS20, a helper plasmid that provides a type IV secretion system. Marker transmission was ...
The dissemination of multi-resistant bacteria represents an enormous burden on modern healthcare. Plasmid-borne conjugative transfer is the most prevalent mechanism, requiring a type IV secretion system that enables bacteria to spread beneficial traits, such as resistance to last-line antibiotics, among different genera. Inc18 plasmids, like the Gram-positive broad host-range plasmid pIP501, are substantially involved in propagation of vancomycin resistance from Enterococci to methicillin-resistant strains of Staphylococcus aureus. Here, we identified the small cytosolic protein TraN as a repressor of the pIP501-encoded conjugative transfer system, since deletion of traN resulted in upregulation of transfer factors, leading to highly enhanced conjugative transfer. Furthermore, we report the complex structure of TraN with DNA and define the exact sequence of its binding motif. Targeting this protein-DNA interaction might represent a novel therapeutic approach against the spreading of antibiotic ...
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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 ...
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 ...
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SUMMARY: Transfer of RP4 and related replicons belonging to the Escherichia coli incompatibility group P (Pseudomonas aeruginosa IncP1) to races 2 and 6 of P. syringae pv. pisi was associated with the creation of two types of transconjugant, one resembling the parental race and the other showing an altered cultivar-specificity towards pea. The latter, irrespective of the parental race, exhibited a novel pattern of interaction with pea that corresponded to race 4; consequently such transconjugants were termed race 4-like. Curing of RP4 did not affect the phenotype, except in relation to the antibiotic resistances specified by RP4. The race 4-like strains were non-fluorescent when cultured on appropriate media (in contrast to the particular isolates of races 2 and 6 from which they were derived), showed an enhanced ability to inherit RP4 subsequently (at frequencies up to 10-1 per recipient) and differed from their parental race in their pattern of plasmid profile. The plasmid profiles were similar for
We develop a system for implementing "packet-based" intercellular communication in an engineered bacterial population via conjugation. 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. Integrase-mediated editing of the address on the message plasmid allows cells to dynamically update the messages recipients in vivo. As a proof-of-concept demonstration of our system, we propose a linear path scheme that would propagate a message sequentially through the strains of a population in a defined order ...
Integrative and conjugative elements (ICEs) are a diverse group of mobile genetic elements found in both Gram-positive and Gram-negative bacteria. ICEs are self-transmissible elements that encode a full complement of machinery for conjugation as well as intricate regulatory systems to control excision from the chromosome and onward conjugative transfer [Wozniak and Waldor, 2010; Burrus,2004]. These multi-talented entities can promote their own mobilization and potentially that of other hitch-hiking genetic elements and thus contribute to horizontal transfer of virulence determinants, antibiotic-resistance genes and other bacterial traits [Hastings. et al., 2004]. ICEs are being identified in increasing numbers as sequenced genome databases expand exponentially [Wozniak, et al., 2010; Ryan, et al., 2009; te Poele, et al., 2008; Burrus et al., 2002]. At present only a few have been classified into ICE families, amongst the best characterized of which is the SXT/R391 family of Vibrio cholerae, ...
p>The checksum is a form of redundancy check that is calculated from the sequence. It is useful for tracking sequence updates.,/p> ,p>It should be noted that while, in theory, two different sequences could have the same checksum value, the likelihood that this would happen is extremely low.,/p> ,p>However UniProtKB may contain entries with identical sequences in case of multiple genes (paralogs).,/p> ,p>The checksum is computed as the sequence 64-bit Cyclic Redundancy Check value (CRC64) using the generator polynomial: x,sup>64,/sup> + x,sup>4,/sup> + x,sup>3,/sup> + x + 1. The algorithm is described in the ISO 3309 standard. ,/p> ,p class="publication">Press W.H., Flannery B.P., Teukolsky S.A. and Vetterling W.T.,br /> ,strong>Cyclic redundancy and other checksums,/strong>,br /> ,a href="http://www.nrbook.com/b/bookcpdf.php">Numerical recipes in C 2nd ed., pp896-902, Cambridge University Press (1993),/a>),/p> Checksum:i ...
Link-A-Light ATTO700 Conjugation Kit, 1 kit. The Link-A-Light conjugation kit allows Fluorescent conjugations to set up in |em|seconds|/em|, simply by adding a solution of the antibody to be labeled to a proprietary lyophilised mixture containing a
Link-A-Light ATTO633 Conjugation Kit, 1 kit. The Link-A-Light conjugation kit allows Fluorescent conjugations to set up in |em|seconds|/em|, simply by adding a solution of the antibody to be labeled to a proprietary lyophilised mixture containing a
Cy3 conjugation / labeling in | 4 hrs with 30 secs hands-on time using Lightning-Link® Cy3 Conjugation Kit (Fast) ab188287. 100% antibody recovery.
The discovery of the process of conjugation in prokaryotes was due to one of the most fortuitous experimental designs in recent scientific history. Many other scientists had tried to demonstrate...
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抗山羊(ab6697)经WB, IP, ELISA, IHC-P, IHC-Fr, ID, Conjugation, ICC/IF实验严格验证。未偶联形式。其他多种未偶联二抗可供选择。品质保证,中国80%以上现货。
抗鼠未偶联(ab7056)经WB, IP, ELISA, IHC-P, IHC-Fr, ID, Conjugation, ICC/IF实验严格验证。产品经预吸附处理(去除交叉反应)。其他多种未偶联二抗可供选择。
Bacterial conjugation is the transfer of genetic material between bacterial cells by direct cell-to-cell contact or by a bridge-like connection between two cells.[1] Conjugation is a mechanism of horizontal gene transfer, as are transformation and transduction, though these two other mechanisms do not involve cell-to-cell contact.[2] Bacterial conjugation was discovered by Nobel Prize winners Joshua Lederberg and Edward Tatum. They showed that the bacterium Escherichia coli entered a sexual phase during which it could share genetic information.[3] Bacterial conjugation is often incorrectly regarded as the equivalent of sexual reproduction, since it involves the exchange of genetic material. During conjugation the donor cell provides a conjugative or mobilizable genetic element that is most often a plasmid or transposon.[4][5] Most conjugative plasmids have systems ensuring that the recipient cell does not already contain a similar element. The genetic information transferred is often beneficial ...
The Gram-positive pathogen Clostridium perfringens possesses a family of large conjugative plasmids that is typified by the tetracycline resistance plasmid pCW3. Since these plasmids may carry antibiotic resistance genes or genes encoding extracellular or sporulation-associated toxins, the conjugative transfer of these plasmids appears to be important for the epidemiology of C. perfringens-mediated diseases. Sequence analysis of members of this plasmid family identified a highly conserved 35kb region that encodes proteins with various functions, including plasmid replication and partitioning. The tcp conjugation locus also was identified in this region, initially based on low-level amino acid sequence identity to conjugation proteins from the integrative conjugative element Tn916. Genetic studies confirmed that the tcp locus is required for conjugative transfer and combined with biochemical and structural analyses have led to the development of a functional model of the Tcp conjugation ...
Define conjugant. conjugant synonyms, conjugant pronunciation, conjugant translation, English dictionary definition of conjugant. n. Either of a pair of organisms, cells, or gametes undergoing conjugation. n either of a pair of organisms or gametes undergoing conjugation n.
In the majority of cases, the surface-associated multicellular communities found in a wide variety of natural and pathogenic ecosystems are formed in the presence of multiple diverse species and genetically distinct strains. In recent years, well-controlled in vitro biofilm model systems have revealed a diversity of molecular mechanisms contributing to development and maturation of single-species biofilms. The mechanisms underlying the biofilm development in the presence of these multispecies consortia are expected to involve even higher degrees of complexity; however, our understanding of mixed-species biofilms is hampered by the limited number of model systems that have been applied to date. The goal of this study was to test the capacity of a simple in vitro model to reveal factors contributing to the formation of more complex biofilm communities. The suitability of this approach to high-throughput analyses was demonstrated with a systematic survey of a large collection of E. coli isolates ...
The traH gene of the staphylococcal conjugative plasmid pSK41 has recently been shown to encode a lipoprotein (N. Firth, K. P. Ridgway, M. E. Byrne, P. D. Fink, L. Johnson, I. T. Paulsen, and R. A. Skurray, Gene 136:13-25, 1993). Here we report that traH encodes a product recognized as a pheromone by Enterococcus faecalis cells harboring the conjugative plasmid pAD1. The mature traH product is not essential for this phenomenon, as expression of pheromone-like activity was found to require sequences encoding only the pro-TraH signal peptide. ...
Bacterial conjugation systems are highly promiscuous macromolecular transfer systems that impact human health significantly. In clinical settings, conjugation is exceptionally problematic, leading to the rapid dissemination of antibiotic resistance genes and other virulence traits among bacterial populations. Recent work has shown that several pathogens of plants and mammals - Agrobacterium tumefaciens, Bordetella pertussis, Helicobacter pylori and Legionella pneumophila - have evolved secretion pathways ancestrally related to conjugation systems for the purpose of delivering effector molecules to eukaryotic target cells. Each of these systems exports distinct DNA or protein substrates to effect a myriad of changes in host cell physiology during infection. Collectively, secretion pathways ancestrally related to bacterial conjugation systems are now referred to as the type IV secretion family. The list of putative type IV family members is increasing rapidly, suggesting that macromolecular ...
Objectives Antibiotic-polluted environments may function as reservoirs for novel resistance plasmids not yet encountered in pathogens. The aims of this study were to assess the potential of resistance transfer between bacteria from such environments and Escherichia coli, and to characterize the conjugative elements involved. Methods Sediment samples from Kazipally lake and Asanikunta tank, two Indian lakes with a history of severe pollution with fluoroquinolones, were investigated. Proportions of resistant bacteria were determined by selective cultivation, while horizontal gene transfer was studied using a GFP-tagged E. coli as recipient. Retrieved transconjugants were tested for susceptibility by Etest® and captured conjugative resistance elements were characterized by WGS. Results The polluted lakes harboured considerably higher proportions of ciprofloxacin-resistant and sulfamethoxazole-resistant bacteria than did other Indian and Swedish lakes included for comparison (52% versus 2% and 60% versus 7
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.. ...
Publishers Accepted Manuscript: Proper accounting of mass transfer resistances in forward osmosis: Improving the accuracy of model predictions of structural parameter ...
This program is dedicated to the understanding of the forces and mechanisms that have forged the genomic architecture of proteobacteria associated with plants (specifically Rhizobium sp.), both in the short term and with an evolutionary perspective.. The long-term goal is to use this knowledge to develop new strategies in genomic engineering. To this end, our research seeks to understand the mechanisms and evolutionary consequences of homologous recombination, site-specific recombination during plasmid conjugation, regulation of conjugative transfer, functional genomics of plasmids (including functional analysis of genes encoded on plasmids and the role of extracytoplasmic sigma factors in stress responses), as well as molecular systematics, microevolution and phylogeography. Part of this knowledge is being integrated into the generation of new approaches for the generation of programmed deletions and global mutagenesis of the genome.. Our work combines bioinformatic analysis with genetic and ...
DNA transfer across membranes and between cells by conjugation is a clear example of a rapid and natural way to acquire new genetic information, not only between bacteria, but also between yeast, plants and even animal cells. All conjugative systems contain a key protein in the membrane to carry out this process: the DNA transporter. In our system, the DNA transporter is TrwB and its crystallographic structure has been recently solved. The strong structural similarity between TrwB and other well known molecular motors, such as the ATP synthase or ring helicases, suggests that TrwB operates as a motor driving a DNA strand through the transport pore, using the energy derived from ATP hydrolysis. TrwB is the best model in a novel group of molecular motors involved in ssDNA transport across membranes; another example of biological molecular motors that convert chemical energy into mechanical work. To analyze the activity of TrwB, we are going to apply an emerging technique: nanotechnology, which ...
Transformation is a mechanism of genetic transfer between bacteria in which the donor DNAexists cell-free in the recipient bacteriums immediate environment
Bacteria are everywhere simply because they can colonize and adapt to different ecological niches in a very short-term period. One important molecular mechanism underlying the abilities of bacteria to colonize new niches is the acquisition of novel traits by conjugative DNA transfer. Under these circumstances, the so-called variable genome (as opposed to the core genome), which encodes an array of accessory functions (such as antibiotic resistance, specific degradation pathways, symbiosis, and virulence, to name a few), is freely exchanged among bacteria ( 1 ). These newly acquired DNA pieces are represented by intra- or extrachromosomal elements, which may or may not have self-replication and/or auto-transferable capacities. However, all of them participate in the fitness of the bacteria to colonize and to adapt to new niches; thus, they contribute to create new evolutionary patterns ( 2 ). Mobile genetic elements (MGEs) constitute a reservoir of DNA that is shared among bacterial species ( 3 ), and
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 ...
Annales de Dermatologie et de Vénéréologie - Vol. 127 - N° 2 - p. 201 - Iconography : Mucinose cutanée primitive diffuse associée à une anémie réfractaire avec excès de blastes - EM|consulte
View Notes - PS10 from PCB 3063 at University of Florida. -3 to 10-5 . Another 7 do not revert spontaneously at detectable frequency, but can be induced to revert at frequencies ranging from 10-5 to
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Red Line, data from 50 samples completed by Gorovsky and Miao lab, and 10 conjugation samples completed by Pearlman lab; all time points (three GROWTH time points, seven STARVATION time points and ten CONJUGATION time points), triplicates. NOTE: For growing cells, L-l, L-m and L-h correspond respectively to ~1X105 cells/ml, ~3.5X105cells/ml and ~1*106 cells/ml. For starvation, ~2X105 cells/ml were collected at 0, 3, 6, 9, 12, 15 and 24 hours) referred to as S-0, S-3, S-6, S-9, S-12, S-15 and S-24). For conjugation, equal volumes of B2086 and CU428 cells were mixed, and samples were collected at 0, 2, 4, 6, 8, 10, 12, 14, 16 and 18 hours after mixing (referred to as C-0, C-2, C-4, C-6, C-8, C-10, C-12, C-14, C-16 and C-18). Blue and red lines represent the expression values normalized by two differenet methods.. ...
Red Line, data from 50 samples completed by Gorovsky and Miao lab, and 10 conjugation samples completed by Pearlman lab; all time points (three GROWTH time points, seven STARVATION time points and ten CONJUGATION time points), triplicates. NOTE: For growing cells, L-l, L-m and L-h correspond respectively to ~1X105 cells/ml, ~3.5X105cells/ml and ~1*106 cells/ml. For starvation, ~2X105 cells/ml were collected at 0, 3, 6, 9, 12, 15 and 24 hours) referred to as S-0, S-3, S-6, S-9, S-12, S-15 and S-24). For conjugation, equal volumes of B2086 and CU428 cells were mixed, and samples were collected at 0, 2, 4, 6, 8, 10, 12, 14, 16 and 18 hours after mixing (referred to as C-0, C-2, C-4, C-6, C-8, C-10, C-12, C-14, C-16 and C-18). Blue and red lines represent the expression values normalized by two differenet methods.. ...
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A plasmid is a small DNA molecule that is physically separate from, and can replicate independently of, chromosomal DNA within a cell. Plasmids are commonly used to multiply (make many copies of) …
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1 AGGGGTTTTT TGCTGAAAGG AGGAACTATA TCCGGATAAC TACGTCAGGT 51 GGCACTTTTC GGGGAAATGT GCGCGGAACC CCTATTTGTT TATTTTTCTA 101 AATACATTCA AATATGTATC CGCTCATGAG ACAATAACCC TGATAAATGC 151 TTCAATAATA TTGAAAAAGG AAGAGTATGA GTATTCAACA TTTCCGTGTC 201 GCCCTTATTC CCTTTTTTGC GGCATTTTGC CTTCCTGTTT TTGCTCACCC 251 AGAAACGCTG GTGAAAGTAA AAGATGCTGA AGATCAGTTG GGTGCACGAG 301 TGGGTTACAT CGAACTGGAT CTCAACAGCG GTAAGATCCT TGAGAGTTTT 351 CGCCCCGAAG AACGTTCTCC AATGATGAGC ACTTTTAAAG TTCTGCTATG 401 TGGCGCGGTA TTATCCCGTG TTGACGCCGG GCAAGAGCAA CTCGGTCGCC 451 GCATACACTA TTCTCAGAAT GACTTGGTTG AGTACTCACC AGTCACAGAA 501 AAGCATCTTA CGGATGGCAT GACAGTAAGA GAATTATGCA GTGCTGCCAT 551 AACCATGAGT GATAACACTG CGGCCAACTT ACTTCTGACA ACGATCGGAG 601 GACCGAAGGA GCTAACCGCT TTTTTGCACA ACATGGGGGA TCATGTAACT 651 CGCCTTGATC GTTGGGAACC GGAGCTGAAT GAAGCCATAC CAAACGACGA 701 GCGTGACACC ACGATGCCTG TAGCAATGGC AACAACGTTG CGCAAACTAT 751 TAACTGGCGA ACTACTTACT CTAGCTTCCC GGCAACAATT AATAGACTGG 801 ATGGAGGCGG ATAAAGTTGC AGGACCACTT CTGCGCTCGG CCCTTCCGGC 851 ...
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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 ...