After mobilization of pR6K:: The band corresponding to the insertion fragment was present in all of them.
The horizontal gene pool: In particular, plasmids shown in Fig 1 encode two conjugative oriTs. A Scheme of the recombination substrate plasmids. Introduction Universal cloning vectors offer a series of advantages to facilitate their manipulation, which include their small size, presence of multiple cloning sites, selection markers, and high copy number.
The most plausible explanation is that the DNA segment cloned in these plasmids could either confer instability to the cell, or affect plasmid stability, leading to low levels of CAT. TrwB KA affects transfer of both DNA and protein during bacterial conjugation [ 20 ], and a recent study has determined that it affects the ATPase activity of the protein [ 28 ]; Larrea et al.
However, the authors observed sometimes an amplicon of 2 kb, suggesting again IS10 integration in a pSU-derived vector, so we decided to characterize these insertion events. We routinely use the pSU8 family of vectors in E. If you have questions about side effects, contact your health care provider. The reason why certain TrwB variants and the presence of two oriTs in the same molecule could be deleterious to the plasmid or to the cell, remains to be determined.
No such increase was ever observed in the vector alone or in many other constructs using the pSU vector backbone. These vectors are also Cm R and carry the same cat gene as the pSU8 family. Halling SM, Kleckner N.
Chloromycetin is used for treating serious infections caused by certain bacteria. Site-specific integration of foreign DNA into minimal bacterial and human target sequences mediated by a conjugative relaxase. This phenomenon, which may be overcome by the use of mild selection conditions, should be taken into account by the many researchers worldwide using this popular family of cloning vectors.
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The open microbiology journal. However, there are previous works that report insertion in target sequences out of the consensus, and in fact, Kovarik and co-workers redefined the IS10 consensus site to 5'-NPuCNN-NGPyN-3' after analysis of a large collection of IS10 transposition events.
Bacterial plasmids and gene spread. Plasmid pSU remained in its original size at the end of the assay, independently of the Cm concentration applied.
At the end of the experiment, plasmid DNA was extracted and aliquots were run on an agarose gel to compare sizes Fig 3. Three promoters near the termini of IS September 1, ; Published: Although transposon Tn10 is not present in the originally sequenced genome of E.
The target site reported here matches with this newly defined consensus. TrwC-mediated site-specific recombination is controlled by host factors altering local DNA topology. Amplification of the tetracycline resistance determinant of pAMalpha1 in Enterococcus faecalis requires a site-specific recombination event involving relaxase.