Comparison of tellurite resistance determinants from the IncP alpha plasmid RP4Ter and the IncHII plasmid pHH1508a

Abstract

The tellurite resistance (Ter) determinants of the IncHII plasmid pHH1508a and the broad host range IncP alpha plasmid RP4Ter were cloned into pUC8, creating plasmids pDT1364 and pDT1558, respectively. The Ter region of pDT1364 was localized to a 1.25-kilobase region by using Tn1000 insertion mutagenesis. Insertions of Tn1000 into pDT1558 which resulted in tellurite sensitivity spanned 1.75 kilobases of DNA. No similarity between the restriction maps of these two plasmids was observed, and no homology could be detected by DNA-DNA hybridization. Expression in an in vitro transcription-translation system showed that pDT1364 encoded two polypeptides with molecular masses of 23 and 12 kilodaltons (kDa) which were not expressed by pUC8. Some of the Tn1000 insertion mutants did not express the 23-kDa protein. pDT1558 encoded a 40-kDa polypeptide which was not expressed by pUC8. Both Ter determinants were expressed constitutively. Our findings suggest that the mechanisms of Ter encoded by these two plasmids are different.