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PCR-based identification of Vibrio cholerae and the closely related species Vibrio mimicus using the large chromosomal ori sequence of Vibrio cholerae

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Title PCR-based identification of Vibrio cholerae and the closely related species Vibrio mimicus using the large chromosomal ori sequence of Vibrio cholerae
 
Creator Saha, A
Deb, R
Shah, S
Ramamurthy, T
Shinoda, S
Mukhophadyay, AK
Bhadra, RK
 
Subject Microbiology
 
Description The bacterial chromosomal replication origin (ori) sequences are a highly conserved essential genetic element. In this study, the large chromosomal replication origin sequence of Vibrio cholerae (oriCI(VC)) has been targeted for identification of the organism, including the biotypes of serogroup O1. The oriCI(VC) sequence-based PCR assay specifically amplified an 890 bp fragment from all the V. cholerae strains examined. A point mutation in the oriCI(VC) sequence of the classical biotype of O1 serogroup led to the loss of a BglII site, which was utilized for differentiation from El Tor vibrios. Interestingly, the PCR assay amplified a similarly sized ori segment, designated as oriCI(VM), from V. mimicus strains, but failed to produce any amplicon with other strains. Cloning and sequencing of the oriCI(VM) revealed high sequence similarity (96%) with oriCI(VC). The results indicate that V. mimicus is indeed very closely related to V. cholerae. In addition, the BglII restriction fragment length polymorphism (RFLP) between oriCI(VM) and oriCI(VC) sequences allowed us to differentiate the two species. The ori sequence-based PCR-RFLP assay developed in this study appears to be a useful method for rapid identification and differentiation of V. cholerae and V. mimicus strains, as well as for the delineation of classical and El Tor biotypes of V. cholerae O1.
 
Publisher BLACKWELL PUBLISHINGOXFORD9600 GARSINGTON RD, OXFORD OX4 2DQ, OXON, ENGLAND
 
Date 2011-09-20T12:12:19Z
2011-09-20T12:12:19Z
2006
 
Type Article
 
Identifier FEMS MICROBIOLOGY LETTERS
0378-1097
http://hdl.handle.net/123456789/14221
 
Language English