1887

Abstract

Previous reports showed that the large linear plasmid SCP1 of A3(2) contains a 5.4 kb centrally located replication locus. We report here that SCP1 actually contains three internal replication loci. Subcloning of the 5.4 kb sequence identified a 3.2 kb minimal locus (//iteron) that determined propagation in The two newly identified replication genes, and , resembled the gene of circular plasmid pZL12. Transcription start points of the three replication genes were determined. The three replication loci could independently determine propagation in linear mode in . Individual and sequential deletions of the and genes were successful. The SCP1-derived linear plasmids with deletions of the and/or genes still propagated in similar copy numbers, were inherited largely stable and were transferred efficiently by conjugation in . Interestingly, SCP1 can be artificially circularized to yield a 280 kb circular plasmid, circular SCP-1 (C-SCP1), which contains the three replication loci. Strikingly, the copy numbers, inheritance and transfer of C-SCP1 resembled that of the linear SCP1 plasmids. Transcripts of the and genes in linear or artificially circularized SCP1 were detected at all the time points of strain growth.

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2013-10-01
2021-10-17
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