1887

Abstract

Little is known of the transfer and maintenance machinery of the IncP-9 plasmids, which are found in spp. and include both degradative and resistance plasmids. One such plasmid, pM3, which confers resistance to streptomycin and tetracycline, was found repeatedly in species from numerous locations in Belarus. pM3 has a broad host range, but is unable to replicate in enterobacteria at 37 °C and above. A mini derivative, pMT2, was constructed by partial I digestion and ligation with a fragment encoding Km. The complete sequence of pMT2 was determined. Analysis of its 8526 bp of pM3 DNA revealed several ORFs whose predicted polypeptide products were found to have similarity to previously analysed proteins involved in plasmid replication ( gene), transfer (; mating-pair formation gene) and stable maintenance (, genes). The organization of these genes showed similarity to several plasmid systems including the Ti and pSYM plasmids as well as IncP-1 plasmids. Subcloning narrowed down the region required for replication, and identified the putative gene and putative promoter region as able to express incompatibility. deletion mutants were lost from the cell line, and expression of the gene was shown to be controlled by negative autoregulation. A pMT2 derivative with an insertion between the and genes showed very weak, if any, ability to replicate autonomously, suggesting that plasmid maintenance may depend on a close interaction of and functions.

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2000-09-01
2019-12-15
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