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Molecular discrimination and identification of Acetobacter genus based on the partial heat shock protein 60 gene ( hsp60 ) sequences
Author(s) -
Huang ChienHsun,
Chang MuTzu,
Huang Lina,
Chua WenShen
Publication year - 2013
Publication title -
journal of the science of food and agriculture
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.782
H-Index - 142
eISSN - 1097-0010
pISSN - 0022-5142
DOI - 10.1002/jsfa.6231
Subject(s) - biology , acetobacter , phylogenetic tree , genetics , gene , 16s ribosomal rna , polymerase chain reaction , primer (cosmetics) , hsp60 , sequence analysis , genotype , bacteria , heat shock protein , hsp70 , chemistry , organic chemistry
Abstract BACKGROUND To identify the Acetobacter species using phenotypic and genotypic ( 16S rDNA sequence analysis) technique alone is inaccurate. The aim of this study was to use the hsp60 gene as a target for species discrimination in the genus Acetobacter , as well as to develop species‐specific polymerase chain reaction and mini‐sequencing methods for species identification and differentiation . RESULTS The average sequence similarity for the hsp60 gene (89.8%) among type strains was significantly less than that for the 16S rRNA gene (98.0%), and the most Acetobacter species could be clearly distinguished. In addition, a pair of species‐specific primer was designed and used to specifically identify Acetobacter aceti , Acetobacter estunensis and Acetobacter oeni , but none of the other Acetobacter strains. Afterwards, two specific single‐nucleotide polymorphism primers were designed and used to direct differentiate the strains belonging to the species A. aceti by mini‐sequencing assay . CONCLUSION The phylogenetic relationships in the Acetobacter genus can be resolved by using hsp60 gene sequencing, and the species of A. aceti can be differentiated using novel species‐specific PCR combined with the mini‐sequencing technology. © 2013 Society of Chemical Industry