Open AccessComparative genome analysis of L actococcus garvieae using a suppression subtractive hybridization library: discovery of novel DNA signatures
Author(s) -
Kim Wonyong,
Park Hee Kuk,
Thanh Hien Dang,
Lee BoYoung,
Shin Jong Wook,
Shin HyoungShik
Publication year - 2011
Publication title -
fems microbiology letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.899
H-Index - 151
eISSN - 1574-6968
pISSN - 0378-1097
DOI - 10.1111/j.1574-6968.2011.02414.x
Subject(s) - biology , lactococcus , lactococcus lactis , suppression subtractive hybridization , genetics , genomic dna , enterococcus , primer (cosmetics) , microbiology and biotechnology , phylogenetic tree , polymerase chain reaction , genome , gene , bacteria , gene expression , lactic acid , chemistry , cdna library , organic chemistry
L actococcus garvieae , the pathogenic species in the genus L actococcus , is recognized as an emerging pathogen in fish, animals, and humans. Despite the widespread distribution and emerging clinical significance of L . garvieae , little is known about the genomic content of this microorganism. Suppression subtractive hybridization was performed to identify the genomic differences between L . garvieae and L actococcus lactis ssp. lactis , its closest phylogenetic neighbor, and the type species of the genus L actococcus . Twenty‐seven clones were specific to L . garvieae and were highly different from L actococcus lactis in their nucleotide and protein sequences. Lactococcus garvieae primer sets were subsequently designed for two of these clones corresponding to a pyrH gene and a novel DNA signature for application in the specific detection of L . garvieae . The primer specificities were evaluated relative to three previously described 16 S r RNA gene–targeted methods using 32 L actococcus and closely related strains. Both newly designed primer sets were highly specific to L . garvieae and performed better than did the existing primers. Our findings may be useful for developing more stable and accurate tools for the discrimination of L . garvieae from other closely related species.