Construction and Identification of Legionella pneumophila Heat Shock Protein 60 Gene Eukaryotic Expression Vector

Objective To construct and identify the eukaryotic expression vector of the heat shock protein 60(HSP60) gene of Legionella pneumophila.Methods The HSP60 gene of Legionella pneumophila was amplified by PCR and then cloned into vector pUC18 for sequencing.The target gene in the recombinant plasmid(pUC18/Hsp60)was sub-cloned into the eukaryotic expression vector pcDNA3.1(+) and the recombinant pcDNA3.1/HSP60 was constructed.This recombinant plasmid was identified by restriction analysis and PCR.Results Comparison of the sequence of HSP60 gene with that reported in GenBank showed that the identities were 98%.And it was comfirmed that HSP60 gene was successfully inserted into pcDNA3.1.Conclusion We successfully constructed the eukaryotic expression vector pcDNA3.1/HSP60 and it laids a good foundation of the research of new legionellosis vaccine.