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The aim of this study was to construct a DNA vaccine based on the Ag85A/MPT64gene of Mycobacterium tuberculosis (MTB) and analyze its immunogenicity by enzyme-linkedimmunospot (ELISPOT) assay. The fusion gene encoding Ag85A/MPT64 was amplifiedby PCR from the genome of the MTB H37Rv strain and cloned into a eukaryotic expressionvector followed by confirmation using restriction endonuclease digestion and DNAsequencing. The immunogenicity of the recombinant vector was tested in vivo inBALB/c mice. The serum antibody titers against Ag85A/MPT64 were detected by ELISPOTassay. The number of ELISPOT spots for the mice following immunization with Ag85A/MPT64was significantly greater than for the negative and blank controls. A DNA vaccinebased on the gene encoding the Ag85A/MPT64 fusion protein of MTB was successfullyconstructed and expressed. Our data may serve as a foundation for further researchinto the prevention and treatment of tuberculosis and carcinomas.

Construction of a DNA vaccine based on the Mycobacterium tuberculosis Ag85A/MPT64 fusion gene and evaluation of its immunogenicity