Construction of eukaryotic expression vector and DNA immune experiment pcHSP65 of mycobacterium tuberculosis

Objective:To construct nucleotide vaccine based on HSP65 gene of mycobacterium tuberculosis.Methods:The gene encoding heat shock protein 65 kilodalton was amplified from genome of mycobacterium tuberculosis H37Rv strain by polymerase chain reaction(PCR) and correctly inserted into corresponding sites of eukaryotic expression vector pcDNA3.1(-) after restriction endonuclease digestion.The experimental mice was vaccined by this recombinant plasmid DNA.Results:The gene fragment inserted into the vector pcDNA3.1(-) was the HSP65 gene of mycobacterium tuberculosis H37Rv strain,which was confirmed by partial nucleotide sequencing.There was specific antibody produce in experimental mice vaccined by the recombinant plasmid DNA and this could protect mice against infection of mycobacterium tuberculosis.Conclusion:The successful construction of recombinant eukaryotic expression vector based on HSP65 gene of mycobacterium tuberculosis,such recombinant plasmid DNA induced specific antibody production in experimental mice,lays the foundation for further researching in prevention and treatment of tuberculosis.