[Amplification of bacterial heat shock protein 60 gene using inverse PCR method]

A method is presented for rapid in vitro amplification of DNA sequences of bacterial HSP60 gene. In our previous work, using degenerate oligonucleotide primers for conserved regions of HSP60 gene, a 600 bp fragment was amplified by PCR, cloned and sequenced. An inverse PCR method, with the primers oriented in the reversed direction of the usual orientation, is used to amplify the DNA sequences that flank the 600 bp known region. The feasibility of this method is shown by amplifying the complete HSP60 gene sequences of 4 bacterial strains, namely Bifidobacterium denticolens, Bifidobacterium inopinatum, Bifidobacterium adolescentis and Gardnerella vaginalis.