1887

Abstract

Multidrug-resistant staphylococci often harbour plasmids that carry genes conferring resistance to several antimicrobial compounds. Many of these multiresistance plasmids appear to utilize a related theta-type replication system for which multiresistance plasmid pSK1 serves as a prototype. Essential pSK1 replication elements were identified by cloning segments of the replication region and testing the resulting plasmids for replication proficiency. An iterated region within and a DNA segment of up to 68 bp upstream of the promoter were both found to be essential for origin activity. The Rep protein was overexpressed as a 6×His-tagged C-terminal fusion protein and was shown to bind to four Rep boxes located within the coding region. Inactivation of a divergently oriented promoter upstream of , designated P, resulted in an elevated plasmid copy number. Comparative analyses suggest that the replication systems of many staphylococcal multiresistance plasmids share a similar genetic organization and utilize an antisense-RNA-mediated regulatory mechanism for copy number control.

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2008-10-01
2020-04-05
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