Gene cloning and sequence analysis of HSP70 from Vibrio alginolyticus

Objective The objective of this study was to analyze the gene sequence of heat shock protein 70 of Vibrio alginolyticus and compare the gene sequence of heat shock protein 70 among vibrio stains reported in GenBank.MethodsA pair of primers was designed according to the nucleotide sequence of the heat shock protein 70 gene of vibrio reported in GenBank.With the specific primers,target fragments were amplified from Vibrio alginolyticus genomic DNA by PCR.The fragments were cloned and sequenced.Results Results indicated that the gene of HSP70 in Vibrio alginolyticus which shared 90.4%-96.2% homology of nucleotides with other vibrio stains,included an ORF of 1 914 bp and encoded a polypeptide of 637 amino acids that contained the characteristic sequence of Dnak (DLGTT-S-V).The N-terminal amino acid sequence of the entire molecule was more conserved than the C terminal.Conclusion Results showed that the HSP70 isolated from Vibrio alginolyticus was a kind of Dnak that had a high homology of nucleotides with Vibrio harveyi.