Characterization of a Cryptic Rolling-Circle Replication Plasmid pMK8 from Enterococcus durans 1-8

Current Microbiology
Penyu ZhouYanling Hao

Abstract

A novel cryptic plasmid from Enterococcus durans 1-8, designated as pMK8, was sequenced and analyzed in this study. It consists of 3337 bp with a G + C content of 33.11%. Sequence analysis of pMK8 revealed three putative open reading frames (ORFs). Based on homology, two of them were identified as genes encoding replication initiation (RepC) and mobilization (Mob) protein, respectively. Sequence analysis revealed a pT181 family double-strand origin (dso) and a putative single-strand origin (sso) located upstream of the repC gene. Sequence homology analysis indicated that the sso belongs to the ssoW family. Southern hybridization confirmed the presence of single-strand DNA (ssDNA) intermediates, suggesting that pMK8 replicates via the RCR mechanism. Furthermore, the relative copy number of pMK8 was estimated by real-time PCR to be 175 ± 14 copies in each cell.

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Citations

Oct 6, 2020·Frontiers in Microbiology·Nils F HülterTal Dagan

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Datasets Mentioned

BETA
KLDS6.0933

Methods Mentioned

BETA
PCR
electrophoresis

Software Mentioned

CLC Sequence Viewer
Basic Local Alignment Search Tool ( BLAST )
DNAMAN
SWISS
BLAST
MODEL
ClustalX

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