title: Molecular discrimination and identification of Acetobacter genus based on the partial heat shock protein 60 gene (hsp60) sequences.
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.
  
  
  CONCLUSIONS
  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.
authors.affiliation: Bioresource Collection and Research Center, Food Industry Research and Development Institute, 331 Shih-Pin Road, Hsinchu, 30062, Taiwan, ROC.
authors.forename: Chien-Hsun
authors.initials: CH
authors.lastname: Huang
authors.forename: Mu-Tzu
authors.initials: MT
authors.lastname: Chang
authors.forename: Lina
authors.initials: L
authors.lastname: Huang
authors.forename: Wen-Shen
authors.initials: WS
authors.lastname: Chua
mesh_terms: D000091:Acetobacter / Q000235:genetics*; D001483:Base Sequence; D018834:Chaperonin 60 / Q000235:genetics*; D017931:DNA Primers; D004269:DNA, Bacterial; D004275:DNA, Ribosomal; D005798:Genes, Bacterial; D005819:Genetic Markers; D005838:Genotype; D010802:Phylogeny; D016133:Polymerase Chain Reaction / Q000379:methods*; D020641:Polymorphism, Single Nucleotide; D012329:RNA, Bacterial; D012336:RNA, Ribosomal, 16S / Q000235:genetics; D017422:Sequence Analysis, DNA / Q000379:methods; D013045:Species Specificity
chemical_list: D018834:Chaperonin 60; D017931:DNA Primers; D004269:DNA, Bacterial; D004275:DNA, Ribosomal; D005819:Genetic Markers; D012329:RNA, Bacterial; D012336:RNA, Ribosomal, 16S
keywords: Acetobacter; direct sequencing; hsp60 gene; mini-sequencing; species-specific PCR