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  • Title: Characterization of a novel rolling-circle replication plasmid pYSI8 from Lactobacillus sakei YSI8.
    Author: Zhai Z, Hao Y, Yin S, Luan C, Zhang L, Zhao L, Chen D, Wang O, Luo Y.
    Journal: Plasmid; 2009 Jul; 62(1):30-4. PubMed ID: 19275914.
    Abstract:
    A plasmid from Lactobacillus sakei YSI8, designated as pYSI8, was sequenced and characterized. It consisted of a 4973bp circular molecule with a G+C content of 35.6%. The plasmid pYSI8 was predicted to contain five putative ORFs, in which ORF1 shared 79% and 76% identity with Rep proteins of pLH2 and pLC2, members of rolling-circle replication (RCR) pMV158 family. Detection of single-stranded DNA (ssDNA) intermediates by Southern hybridization and mung bean nuclease treatment confirmed that pYSI8 replicated via the RCR mechanism. Accumulation of ssDNA in rifampicin-treated strains implied that the host-encoded RNA polymerase was involved in the conversion of ssDNA to double-stranded DNA (dsDNA). Furthermore, the copy number of pYSI8 was estimated to be 41.9+/-0.5 in each cell by real-time polymerase chain reaction.
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