Abstract
Isolates ofRhodococcus equifrom 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 1.35-kb fragment containing the origin of replication ofR. equistrain 103 virulence-associated plasmid (pOTS) was identified, sequenced, and its location identified. A simpleR. equi–Escherichia colishuttle plasmid (pRE-1) derived from theE. coliplasmid pACYC177 and the pOTSoriwas developed. The plasmid transformed readily and was stable in either host and expressed kanamycin resistance but not β-lactamase inR. equi.An improved 5.9-kb vector, pRE-7, was developed from pRE-1 and pBluescript. Subcloning ofvapAinto the multiple cloning site of the β-galactosidase gene of pRE-7 resulted in weak expression of the gene both inE. coliandR. equi.The shuttle vector may be useful in examining regulation of virulence gene expression inR. equi.
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