Construction and identification of a eukaryotic expression vector of HSP70 from Vibrio alginolyticus

Objective To construct and identify a eukaryotic expression vector pcDNA3.1 of HSP70 from Vibrio alginolyticus.Methods Genomic DNA was extracted from V.alginolyticus.The HSP70 gene was amplified from V.alginolyticus genomic DNA by PCR.The purified PCR fragment was ligated into the pMD18-T simple vector;the positive clones were screened and identified by PCR and restriction enzyme digestion and the positive clones were sequenced after identification.The HSP70 gene fragments from TA clones were cloned into the eukaryotic expression vector pcDNA3.1 using restriction enzymes.The recombinant plasmid pcDNA3.1was identified through PCR,enzyme digestion and sequencing.Results The HSP70 gene fragment was amplified correctly,as the size of gene was a 1 914 bp,and HSP70 eukaryotic expression vector pcDNA3.1 was successfully constructed. Conclusion Construction of the eukaryotic expression vector pcDNA3.1 of HSP70 from V.alginolyticus lays the foundation for the development of HSP70 vaccine.