| Title: | Genetic characterization of the conjugative DNA processing system of enterococcal plasmid pCF10 |
| Author(s): | Staddon JH; Bryan EM; Manias DA; Chen Y; Dunny GM; |
| Address: | "Department of Microbiology, University of Minnesota Medical School, Minneapolis, MN 55455-0312, USA" |
| DOI: | 10.1016/j.plasmid.2006.05.001 |
| ISSN/ISBN: | 0147-619X (Print) 1095-9890 (Electronic) 0147-619X (Linking) |
| Abstract: | "Conjugation is a major contributor to lateral gene transfer in bacteria, and pheromone-inducible conjugation systems in Enterococcus faecalis play an important role in the dissemination of antibiotic resistance and virulence in enterococci and related bacteria. We have genetically dissected the determinants of DNA processing of the enterococcal conjugative plasmid pCF10. Insertional inactivation of a predicted relaxase gene pcfG, via insertion of a splicing-deficient group II intron, severely reduced pCF10 transfer. Restoration of intron splicing ability by genetic complementation restored conjugation. The pCF10 origin of transfer (oriT) was localized to a 40-nucleotide sequence within a non-coding region with sequence similarity to origins of transfer of several other plasmids in gram positive bacteria. Deletion of the oriT reduced pCF10 transfer by more than five orders of magnitude without affecting pCF10-dependent mobilization of co-resident oriT-containing plasmids. Although the host range for pCF10 replication is limited to enterococci, we found that the pCF10 conjugation system promotes mobilization of oriT-containing plasmids to multiple bacterial genera. Therefore, this transfer system may have applications for gene delivery to a variety of poorly-transformed bacteria" |
| Keywords: | "Base Sequence Chromosome Mapping Computational Biology Conjugation, Genetic/*genetics DNA Primers DNA, Bacterial/*genetics Electroporation Enterococcus faecalis/*genetics Gene Transfer Techniques Molecular Sequence Data Plasmids/*genetics Polymerase Chain;" |
| Notes: | "MedlineStaddon, Jack H Bryan, Edward M Manias, Dawn A Chen, Yuqing Dunny, Gary M eng T32 GM008347/GM/NIGMS NIH HHS/ T32GM08244/GM/NIGMS NIH HHS/ 1R01-GM49530/GM/NIGMS NIH HHS/ T32GM08347/GM/NIGMS NIH HHS/ R01 GM049530/GM/NIGMS NIH HHS/ R01 GM049530-20/GM/NIGMS NIH HHS/ T32 GM008244/GM/NIGMS NIH HHS/ T32AI07421/AI/NIAID NIH HHS/ T32 AI007421/AI/NIAID NIH HHS/ Comparative Study Research Support, N.I.H., Extramural 2006/06/16 Plasmid. 2006 Sep; 56(2):102-11. doi: 10.1016/j.plasmid.2006.05.001. Epub 2006 Jun 13" |
