Molecular analysis of oriT and MobA protein in the 7.4 kb mobilizable beta-lactamase plasmid pSJ7.4 from Neisseria gonorrhoeae

Plasmid
Nydia M Rodriguez-Bonano, L J Torres-Bauzá

Abstract

The mobilization region of the 7.4 kb beta-lactamase plasmid pSJ7.4 from Neisseria gonorrhoeae was characterized. The 3.2 kb HindIII-BamHI fragment of pSJ7.4 was mobilized between Escherichia coli strains by conjugative plasmid RK2. Selected restriction enzyme-generated deletions of this fragment were subcloned in pACYC177 to obtain constructs that were suitable for analysis of the mobilization region. Mating experiments showed that a highly conserved 1.9 kb DNA region within coordinates 4096-5997 is required for mobilization of pSJ7.4. This region contains two genes encoding the mobilization protein MobA, another protein, MobC, putatively involved in mobilization from some hosts, and an intergenic oriT. The 168 bp intergenic sequence also contains the promoters for mobA and mobC in an arrangement that suggests divergent transcription and autoregulation from oriT. The 56 kDa MobA was expressed in E. coli as a (6x)His-Tag fusion protein. Purified MobA specifically induced plasmid relaxation by nicking at the oriT. MobA is exceptional because the N-terminal region alone can mobilize pSJ7.4, albeit at a lower frequency than the full-length protein, even in the absence of MobC. The carboxyl terminal region of MobA did not share hom...Continue Reading

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Citations

Jan 27, 2006·Antimicrobial Agents and Chemotherapy·Mónica BlancoJesús Navas
Apr 9, 2010·PloS One·Emilia Pachulec, Chris van der Does
Jun 25, 2017·Pathogens and Disease·Ana Cehovin, Steven B Lewis

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