Construction of Staphylococcus plasmid vector pCA43 conferring resistance to chloramphenicol, arsenate, arsenite and antimony

Abstract

The arsenate (Asa), arsenite (Asi) and antimony (III) (Amo) resistance region of the Staphylococcus xylosus 29.5-kb plasmid pSX267 has been recloned in S. carnosus using the chloramphenicol resistance (CmR) plasmid pC194. In several deletion steps we constructed a 5.9-kb plasmid, pCA43, which confers resistance to Cm, Asa, Asi and Amo salts. pCA43 possesses unique sites for the restriction endonucleases PvuII, StuI, BamHI, AvaII, HindIII, PstI, XbaI and BclI. Insertional inactivation was achieved with StuI (affecting Cm resistance), BamHI (affecting only Asa resistance), AvaII, HindIII and PstI (affecting Asa, Asi and Amo resistances). Plasmid stability was tested and found to be high after DNA insertion into the BamHI or HindIII sites.