Characterization of a Rolling-Circle Replication Plasmid pLR1 from <Emphasis Type="Italic">Lactobacillus plantarum</Emphasis> LR1 |
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Authors: | Ruoyu Li Zhengyuan Zhai Sheng Yin Ying Huang Qing Wang Yunbo Luo Yanling Hao |
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Institution: | (1) College of Food Science and Nutritional Engineering, China Agricultural University, No. 17 Qing Hua East Road, Hai Dian District, Beijing, China |
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Abstract: | A cryptic plasmid from Lactobacillus plantarum LR1, designated pLR1, was sequenced and characterized. It consisted of a 2066-bp circular molecule with a G + C content of
52.7%. The plasmid pLR1 was predicted to contain five putative ORFs, in which ORF1 shared 93% and 92% identity with Rep proteins
of pLP1 and pC30il, members of rolling-circle replication (RCR) pC194 family. Detection of single-stranded DNA (ssDNA) intermediates
by Southern hybridization and mung bean nuclease treatment confirmed that pLR1 replicated via the RCR mechanism. Accumulation
of ssDNA in rifampicin-treated strains implied that the host-coded RNA polymerase was involved in the conversion of ssDNA
to double-stranded DNA (dsDNA). Furthermore, the copy number of pLR1 was estimated to be 36 in each cell by real-time polymerase
chain reaction. |
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