Vectors coli plasmids. Medical search. Frequent questions

Artificial plasmids are widely used as vectors in molecular cloning, serving to drive the replication of recombinant DNA sequences within host organisms. (wikipedia.org)
This technique allows efficient creation of scarless recombinant plasmids at many, but not all, positions in a vector. (neb.com)
These fragments are then inserted into cloning vectors , such as bacterial plasmids or bacteriophages, which transfer the recombinant DNA to suitable host cells, such as the bacterium E. coli. (encyclopedia.com)
It relies on generation of recombinant virus by site-specific transposition in E. coli (DH10Bac™) rather than homologous recombination in insect cells. (fishersci.com)
Antibiotics are used to select all types of plasmidic vectors in bacteria which is a significant concern in industrial cultures producing recombinant proteins or DNA. (igem.org)
As plasmid-free cells grow faster than plasmid-carrying cells, the yield and the production reproducibility of recombinant molecules are significantly lowered in the absence of plasmid selection by the appropriate antibiotic. (igem.org)
Recombinant expression of proteins of interest in Escherichia coli is an important tool in the determination of protein structure. (iucr.org)
The goal of this work was to evaluate the effect of a recombinant beta-carotene 15, 15'-monooxygenase (BCMO1) from Gallus gallus, expressed in Escherichia coli. (bvsalud.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)
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)
This plasmid contains a constitutive bacterial promoter that does not require induction. (sigmaaldrich.com)
We also offer a range of other bacterial promoters that are compatible with this plasmid and are available on request. (sigmaaldrich.com)
The data also showed evidence of full integration of the plasmid into the bacterial chromosome. (oregonstate.edu)
The pLATE bacterial expression vectors are designed for high levels of target protein expression in concert with minimal background (uninduced) expression, which permits expression of proteins that are toxic to E. coli cells. (thermofisher.com)

To download a certificate of analysis for pEN_CmiR-Luc-A Plasmid in Escherichia coli ( MBA-304 ), enter the lot number exactly as it appears on your product label or packing slip. (atcc.org)
The certificate of analysis for that lot of pEN_CmiR-Luc-A Plasmid in Escherichia coli ( MBA-304 ) is not currently available online. (atcc.org)
PCR Primer Design for 16S rRNAs for Experimental Horizontal Gene Transfer Test in Escherichia coli. (redasoft.com)
We just lately demonstrated that the Escherichia coli ribosome is strong sufficient to accommodate international 16S rRNAs from numerous gamma- and betaproteobacteria micro organism (Kitahara et al. (redasoft.com)
A plasmid vector was constructed that allowed the expression of biotinylated scFvs in Escherichia coli. (usda.gov)
1979. Molecular cloning of polyoma virus DNA in escherichia coli: plasmid vector systems. (wisc.edu)
2) pSET152, an Escherichia coli plasmid with ϕC31 attP site, and pHZ1358, a Streptomyces-Escherichia coli shuttle cosmid vector, both carrying oriT from RP4, can be mobilized from E. coli into NS3226 by conjugation. (microbiologyresearch.org)
Plasmid cloning vectors for the conjugal transfer of DNA from Escherichia coli to Streptomyces spp. (microbiologyresearch.org)
In this study, choline dehydrogenase ( betA ) gene encoding for glycine betaine biosynthesis in Escherichia coli isolated from salted shark ( Scoliodon sp. (scialert.net)
Escherichia coli was isolated from salted shark ( Scoliodon sp. (scialert.net)
Escherichia coli JM109 and plasmid pTZ57R/T (MBI Fermentas, Hanover, Maryland, USA) were used as transformation host and cloning vector, respectively. (scialert.net)
In this study, the sp2 gene was cloned and expressed in Escherichia coli to produce a 6xHis-tagged fusion protein, which was then used to generate a polyclonal antibody. (hindawi.com)
The expressed protein in Escherichia coli was used to generate polyclonal antibodies. (hindawi.com)
Escherichia coli strains TG1 and BL21 (DE3) were grown at 37°C in LB medium. (hindawi.com)
An impediment to the successful production of large quantities of natively folded proteins in Escherichia coli is the tendency of many proteins to become insoluble when overexpressed. (iucr.org)
In Escherichia coli , besides the PEPCK reaction, there is another pathway to form PEP from C 4 -dicarboxylic acid which involves the NAD- and NADP-dependent malic enzymes (MAEA and MAEB) and phosphoenolpyruvate synthase (PPS). (asm.org)

This plasmid is designed to express tagged proteins in E. coli. (sigmaaldrich.com)
This plasmid contains an Outer Membrane Protein T (OmpT) secretory signal peptide (SP) to allow proteins to be exported from the cytosol. (sigmaaldrich.com)
For very toxic proteins, we recommend using the pQE-80L series of expression vectors in the M15[pREP4] E. coli host strain . (qiagen.com)
Those include its use as a critical starting material for transient transfection to produce both viral-vector constructs (e.g., lentivirus or adenoassociated virus) and transient proteins in development (e.g., monoclonal antibody). (bioprocessintl.com)
The pFastBac™ HBM TOPO™ vector has also a C-terminal His-Tag with a TEV cleavage signal to enable easy purification of Histidine fusion proteins on nickel-chelating resins (ProBond™ Purification System) and native proteins with the aid of AcTev™ protease. (fishersci.com)
If a plasmid carrying the ccd module is lost at cell division, the plasmid-free bacteria contain poison and antidote proteins in their cytoplasm. (igem.org)

We explored the utility of a broad host-range plasmid, pBBR1MCS-4, to transform chlamydiae, with a goal of simplifying the transformation process. (oregonstate.edu)
In theStaby TM system,the antidote gene (ccdA) is introduced in the plasmid DNA under the control of a weak constitutive promoter: the mob gene promoter, which originates from a broad host range plasmid (pBHR1). (igem.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)
The plasmid contains a constitutive promoter (OXB20) derived from the region upstream of the E. coli RecA gene. (sigmaaldrich.com)
The vectors in the cis-Repressed pQE Vector Set are similar to those in the N-Terminus pQE Vector Set, but additionally express the laclq gene product that represses protein expression prior to IPTG induction. (qiagen.com)
This eliminates the need to include this gene on another plasmid. (qiagen.com)
Use of these vectors is recommended when expressing gene products toxic to E. coli. (qiagen.com)
The cis-Repressed pQE Kan Vector Set contains vectors with a kanamycin resistance gene in addition to the ampicillin resistance gene providing selection by either antibiotic. (qiagen.com)
The pQE-80L vectors have a cis- lacIq gene that overexpresses the lac repressor, in addition to a lacI repressor gene present in trans on a separate pREP4 plasmid. (qiagen.com)
Note that times are based on estimates for moving a gene from one plasmid to another. (neb.com)
Generated plasmids have been used in clinical trials for gene therapy and vaccines as well as in viral vector production. (bioprocessintl.com)
Thermo Scientific aLICator™ LIC Cloning and Expression System is designed for fast and efficient ligation independent cloning and tight regulation of gene expression in E. coli . (thermofisher.com)
The tightly regulated expression and fast, efficient directional cloning makes the aLICator LIC Cloning and Expression System the best choice for routine and toxic gene cloning and expression in E. coli . (thermofisher.com)
On the other hand, the toxic gene (ccdB) is introduced in the E. coli chromosome of the bacteria. (igem.org)
Expression of the poison gene is under the control of a promoter strongly repressed in the presence of the plasmid. (igem.org)
Understanding the presence of plasmid vectors requires an appreciation of the basics of gene-editing, which, confusingly, are considerably distinct from what the word 'editing' means in ordinary English. (greenmedinfo.com)
Does gene orientation relative to origin of replication matter on small plasmids? (stackexchange.com)
How do you read the gene insert domains on a plasmid from Addgene? (stackexchange.com)

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)
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)
PCR and PacBio-based whole genome sequencing were used to determine if the plasmid was maintained within the chromosome or as an episome. (oregonstate.edu)
PacBio sequencing of the cloned transformants revealed allelic exchange events between the chromosome and plasmid pBVR2 that replaced chromosomal incA with the plasmid GFP:CAT cassette. (oregonstate.edu)

Conjugal Transfer, Whole Genome Sequencing, and Plasmid Analysis of Four mcr-1 -bearing Isolates from U.S. Patients. (cdc.gov)

In nature, plasmids often carry genes that benefit the survival of the organism and confer selective advantage such as antibiotic resistance. (wikipedia.org)
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)
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)
Many of the genes carried by a plasmid are beneficial for the host cells, for example: enabling the host cell to survive in an environment that would otherwise be lethal or restrictive for growth. (wikipedia.org)
To overcome the plasmid instability, many antibiotic resistance genes are used as selectable markers in fermentation processes. (igem.org)

Qualified cell banks should be used and tested for identity, viability, strain, genotype/phenotype, and presence/copy number of the plasmid vector of interest, along with a description of its structural elements. (bioprocessintl.com)
The final plasmid construct, pBVR2, was successfully transformed into C. trachomatis strain L2-434. (oregonstate.edu)
1) Streptomyces phage ϕC31 and its derived vectors can infect and lysogenize this strain. (microbiologyresearch.org)
The plasmids were transformed into an E. coli TOP10 strain and they were plated in agar plates containing Chloramphenicol and Amipiciline. (igem.org)

Plasmids can also provide bacteria with the ability to fix nitrogen. (wikipedia.org)
Practically, 100 % of the bacteria will carry the vector. (igem.org)
Can bacteria pick up lethal plasmids? (stackexchange.com)
To be able to replicate a plasmid in bacteria, you need a origin of replication (ori). (stackexchange.com)

The plasmid was modified to contain chromosomal DNA from C. trachomatis to facilitate homologous recombination. (oregonstate.edu)

pQE vectors provide high-level expression of protein in E. coli and add the small 6xHis affinity tag to protein expression constructs. (qiagen.com)
Once a protein-encoding DNA insert has been incorporated into the vector, it is transformed into E. coli cells and protein expression is induced by the addition of IPTG to the growth medium. (qiagen.com)
The new pFastBac™ HBM TOPO™ vector enables secreted protein expression because it has the honeybee melittin (HBM) secretion signal. (fishersci.com)
aLICator™ LIC Cloning and Expression Kit 1 - pLATE11 vector, untagged protein expression. (thermofisher.com)
aLICator™ LIC Cloning and Expression Kit 2 (N-terminal His-tag/EK) -pLATE51 vector, N-terminal His-tag protein expression. (thermofisher.com)
aLICator™ LIC Cloning and Expression Kit 3 (C-terminal His-tag) -pLATE31 vector, C-terminal His-tag protein expression. (thermofisher.com)
aLICator™ LIC Cloning and Expression Kit 4 (N-terminal His-tag/WQ) -pLATE 52 vector, N-terminal His-tag protein expression, WELQut cleavage. (thermofisher.com)
aLICator™ LIC Cloning and Expression Set 1 (All-in-One/EK) -pLATE11, pLATE51 and pLATE31 vectors, choice of untagged, N- or C-terminal His-tag protein expression. (thermofisher.com)

This BioBricks were into the vectors PSB1C3 y PSB1A2, with resistence to Chloramphenicol and Ampiciline respectively. (igem.org)

However, when pDNA is used as a transfection material, multiple plasmid constructs often are required to produce an individual product coupled with an escalating demand for plasmid quantity as production scales increase. (bioprocessintl.com)

In addition, pDNA sizes used for viral constructs are usually larger (≥11 kb) ( 1 ) and can contain base-pair sequences (such as inverted terminal repeats) that cause problems in their plasmid production. (bioprocessintl.com)

In the laboratory, plasmids may be introduced into a cell via transformation. (wikipedia.org)
Joined fragments have 4 nicks that are repaired by E.coli during transformation. (neb.com)
The product contains 4 nicks, like the original LIC product, and is repaired by E.coli during transformation. (neb.com)
Plasmid transformation requires a replicon derived from the native Chlamydia plasmid, and these transformations are species-specific. (oregonstate.edu)

A plasmid is a small, extrachromosomal DNA molecule within a cell that is physically separated from chromosomal DNA and can replicate independently. (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)
These linear plasmids require specialized mechanisms to replicate their ends. (wikipedia.org)

While some plasmids were fully integrated, some were maintained as episomes and could be purified and retransformed into E. coli. (oregonstate.edu)

The term plasmid was introduced in 1952 by the American molecular biologist Joshua Lederberg to refer to "any extrachromosomal hereditary determinant. (wikipedia.org)
Find out more at Oxford Genetics - Sigma's partner for cloning and expression vectors for molecular biology and synthetic biology applications. (sigmaaldrich.com)

Synthetic plasmids are available for procurement over the internet. (wikipedia.org)

Sequences flanking incA were cloned into the pBBR1MCS-4 vector along with the GFP:CAT cassette from the pSW2-GFP chlamydial shuttle vector. (oregonstate.edu)

Those include an antibiotic-free plasmid maintenance system, operator repressor titration (ORT), and cell-lysis technologies ( 2 - 4 ). (bioprocessintl.com)

Plasmids are generally circular, but examples of linear plasmids are also known. (wikipedia.org)
Covalent bond formation at the vector-insert junctions occurs within the cell to yield circular plasmid. (thermofisher.com)
Is circular DNA the same as plasmids? (stackexchange.com)

This modification of the protocol allows a scarless and sequence-independent insertion into most any vector to be made. (neb.com)

The vector uses the strong polyhedrin promoter to generate high levels of expression in a variety of insect cell lines such as Sf9, Mimic™ Sf9, Sf21, and High Five™ cells in Sf-900 II & Sf-900™ III media from Gibco. (fishersci.com)

Bac-to-Bac™'s pFastBac™ vectors, however, recombines with the parent bacmid in DH10Bac™ E. coli competent cells to form an expression bacmid. (fishersci.com)
pFastBac⁄HBM TOPO™ Vector containing the C-terminal TEV cleavage sit and His-Tag. (fishersci.com)
The pET-28a(+) expression vector and pFastBac HTB vector were conserved in our laboratory. (hindawi.com)

For reference most cloning programs will import a .gb (Genbank) file and will show all of the plasmids features automatically when downloaded and imported. (sigmaaldrich.com)
This method allows rapid creation of compatible ends and avoids the clean-up steps prior to vector and insert joining common to many successful cloning methods. (neb.com)
Garvin L, Vande Voorde R, Dickinson M, Carrell S, Hybiske K, Rockey D (2021) A broad-spectrum cloning vector that exists as both an integrated element and a free plasmid in Chlamydia trachomatis. (oregonstate.edu)
To streamline and facilitate the process of insert cloning into the expression vector, the aLICator system uses directional LIC cloning technology, a rapid procedure that provides high-cloning efficiencies. (thermofisher.com)
The aLICator LIC cloning system uses directional LIC cloning technology to streamline and facilitate cloning into an expression vector. (thermofisher.com)

This allows the exonuclease activity of T4 DNA Polymerase to proceed and generate the complementary overlaps between insert and vector. (neb.com)
One of these interesting approaches is the use of nicking DNA endonucleases (NiDE) to create the complementary overlaps used to anneal the vector and insert. (neb.com)
Alternatively, complementary DNA is inserted into the vectors, or 'naked' DNA fragments can be taken up directly by a host bacterium from its medium (this is less efficient than vector transfer). (encyclopedia.com)

This creative technique uses the 3' → 5' exo activity of T4 DNA Polymerase to create overhangs with complementarity between the vector and insert. (neb.com)
The LIC method uses T4 DNA polymerase to create specific 14 to 21 nucleotide single-stranded overhangs on the pLATE vectors and DNA inserts. (thermofisher.com)

What is the origin of replication and the plasmid copy number of the pQE vectors? (qiagen.com)
The pQE-TriSystem Vector has a pUC origin of replication and is classified as a high-copy vector. (qiagen.com)
Thus, the plasmid can be successfully transformed into chlamydia without a chlamydial origin of replication and can exist in multiple states within a transformed population. (oregonstate.edu)

Plasmids are transmitted from one bacterium to another (even of another species) mostly through conjugation. (wikipedia.org)

After annealing, the LIC vector and insert are transformed into competent E. coli cells without the use of T4 DNA ligase. (thermofisher.com)

Plasmids are considered replicons, units of DNA capable of replicating autonomously within a suitable host. (wikipedia.org)

Some plasmids, however, have no observable effect on the phenotype of the host cell or its benefit to the host cells cannot be determined, and these plasmids are called cryptic plasmids. (wikipedia.org)

When the protein structure and features are not well known, it is recommended to clone into all three vectors and determine the most compatible vector for further research. (thermofisher.com)

This tetrazole compound was used to develop a selective screen by exposing E. coli hosts to the compound and exposing the cells to UV light to induce the photoclick reaction of the tetrazole compound binding to the terminal alkene on botryococcene. (plu.edu)
SOS system in BS to make cells fillamentous (homologue of E.coli solA). (igem.org)
These two biobricks are the main Activator-Inhibitor system, as showned below they are in two different plasmids, and the idea is transform Top10 cells with them in petri dishes with IPTG and ATC. (igem.org)

Inserts are usually PCR amplified and vectors are made linear either by restriction enzyme digestion or by PCR. (neb.com)

Plasmids vary in size from 1 to over 400 kbp, and the number of identical plasmids in a single cell can range anywhere from one to thousands under some circumstances. (wikipedia.org)
Plasmids may be present in an individual cell in varying number, ranging from one to several hundreds. (wikipedia.org)
The normal number of copies of plasmid that may be found in a single cell is called the plasmid copy number, and is determined by how the replication initiation is regulated and the size of the molecule. (wikipedia.org)

Therefore, higher plasmid stability means higher protein of interest production. (igem.org)

The specific enzymes that form botryococcene are SSL-1 and SSL-3 which were expressed in E. coli hosts for this study. (plu.edu)

Their size can range from very small mini-plasmids of less than 1-kilobase pairs (kbp) to very large megaplasmids of several megabase pairs (Mbp). (wikipedia.org)
Plasmids used as therapeutic agents tend to be relatively small in size (4.5- 6.0 kb and even down to 2.5 kb) ( 1 ), requiring relatively low quantities to support early phase clinical trials (1-5 g of purified pDNA). (bioprocessintl.com)

Please see also FAQ 350 for general information on replication origins and copy numbers of various commonly used plasmids. (qiagen.com)

But over the past 24 months, the overall demand and therapeutic applications for plasmid DNA (pDNA) have rapidly grown and expanded. (bioprocessintl.com)

In E . coli , the biosynthetic pathway for the production of glycine betaine from choline has been well characterized at the genetic level ( Landfald and Strom, 1986 ). (scialert.net)

however, some classes of plasmids encode the conjugative "sex" pilus necessary for their own transfer. (wikipedia.org)

The origin of this foreign DNA, as with Recombinetics' calves, was the plasmid vector used in the editing process. (greenmedinfo.com)

The new vector should be strongly considered for the expression of glycoproteins. (fishersci.com)

How to replace intron region in a plasmid? (stackexchange.com)

Anatomy7

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Phenomena and Processes79

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Proteins6

Broad host-range plasmid2

Gene17

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