1887

Abstract

The conjugative multiple antibiotic resistance plasmid pIP501 can be transferred and stably maintained in a variety of Gram-positive genera, including multicellular , as well as in Gram-negative . The 15 putative pIP501 transfer () genes are organized in an operon-like structure terminating in a strong transcriptional terminator. This paper reports co-transcription of the pIP501 genes in exponentially growing JH2-2 cells, as shown by RT-PCR. The genes are expressed throughout the life cycle of , and the expression level is independent of the growth phase. Electrophoretic mobility shift assays indicated that the TraA relaxase, the first gene of the operon, binds to the promoter P, which partially overlaps with the origin of transfer (). DNase I footprinting experiments further delimited the TraA binding region and defined the nucleotides bound by TraA. -Galactosidase assays with P fusions proved P promoter activity, which was strongly repressed when TraA was supplied . Thus, it is concluded that the pIP501 operon is negatively autoregulated at the transcriptional level by the conjugative DNA relaxase TraA.

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2006-03-01
2019-10-22
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