1887

Abstract

A 2019 bp DNA fragment containing the replicon of pSW800 from SW2 was cloned and characterized. This replicon contains two genes – and , which encode a 365 kDa replication initiation protein (RepA) and a peptide of 18 aa, respectively. These two genes overlap by 8 bases with situated upstream. The replicon also transcribes an antisense RNA (RNAI) that inhibits the expression of and . The ribosome-binding sequence (RBS) of is likely to be hidden in a stem–loop structure, inhibiting the translation of . Furthermore, translation of is likely to disrupt the stem–loop structure, which is one of the criteria allowing the translation of to begin. A mutagenesis study revealed that a sequence (5′-GCACGGG-3′) located 111 nt upstream from is crucial; mutation of this sequence prevented the translation of . Additionally, this region and the stem–loop structure containing the RBS of may form an RNA pseudoknot. Results in this study demonstrate that a mechanism similar to that regulating plasmid replication in the IncB, IncIα and IncL/M groups also regulates pSW800 replication.

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2001-10-01
2020-04-06
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