Development of a Rhodococcus equi-Escherichia coli plasmid shuttle vector

Plasmid
H ZhengJ F Prescott

Abstract

Isolates of Rhodococcus equi from pneumonic foals possess an 85- or 90-kb virulence-associated plasmid. A prominent, thermoregulated surface antigen, VapA, encoded by these plasmids is thought to be important in virulence. A 135-kb fragment containing the origin of replication of R. equi strain 103 virulence-associated plasmid (pOTS) was identified, sequenced, and its location identified. A simple R. equi-Escherichia coli shuttle plasmid (pRE-1) derived from the E. coli plasmid pACYC177 and the pOTS ori was developed. The plasmid transformed readily and was stable in either host and expressed kanamycin resistance but not beta-lactamase in R. equi. An improved 5.9-kb vector, pRE-7, was developed from pRE-1 and pBluescript. Subcloning of vapA into the multiple cloning site of the beta-galactosidase gene of pRE-7 resulted in weak expression of the gene both in E. coli and R. equi. The shuttle vector may be useful in examining regulation of virulence gene expression in R. equi.

References

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Citations

Apr 16, 2004·BMC Microbiology·Philip A LessardAnthony J Sinskey
Feb 16, 2006·Journal of Applied Microbiology·P Veiga-CrespoT G Villa
Sep 19, 2006·Veterinary Microbiology·Yanlong PeiJohn F Prescott
May 21, 2009·FEMS Microbiology Reviews·Kristine von Bargen, Albert Haas
Sep 29, 2004·FEMS Microbiology Reviews·Volker GürtlerRobert Seviour
Dec 26, 2001·FEMS Microbiology Letters·M W Mangan, W G Meijer
Jul 18, 2006·FEMS Microbiology Letters·Meijuan ShenZhongjun Qin
Feb 22, 2005·Antonie van Leeuwenhoek·Michael W ManganWim G Meijer

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