title: Construction of a DNA vaccine based on the Mycobacterium tuberculosis             Ag85A/MPT64 fusion gene and evaluation of its immunogenicity.
abstract: The aim of this study was to construct a DNA vaccine based on the Ag85A/MPT64             gene of Mycobacterium tuberculosis (MTB) and analyze its immunogenicity by enzyme-linked             immunospot (ELISPOT) assay. The fusion gene encoding Ag85A/MPT64 was amplified             by PCR from the genome of the MTB H37Rv strain and cloned into a eukaryotic expression             vector followed by confirmation using restriction endonuclease digestion and DNA             sequencing. The immunogenicity of the recombinant vector was tested in vivo in             BALB/c mice. The serum antibody titers against Ag85A/MPT64 were detected by ELISPOT             assay. The number of ELISPOT spots for the mice following immunization with Ag85A/MPT64             was significantly greater than for the negative and blank controls. A DNA vaccine             based on the gene encoding the Ag85A/MPT64 fusion protein of MTB was successfully             constructed and expressed. Our data may serve as a foundation for further research             into the prevention and treatment of tuberculosis and carcinomas.
authors.affiliation: The Second Hospital of Jilin University, Changchun 130041, P.R.                     China.
authors.forename: Hong
authors.initials: H
authors.lastname: Bao
authors.forename: Ting
authors.initials: T
authors.lastname: Yu
authors.forename: Yufen
authors.initials: Y
authors.lastname: Jin
authors.forename: Chunyan
authors.initials: C
authors.lastname: Teng
authors.forename: Ximing
authors.initials: X
authors.lastname: Liu
authors.forename: Yanlei
authors.initials: Y
authors.lastname: Li
mesh_terms: D000217:Acyltransferases / Q000235:genetics* / Q000378:metabolism; D000818:Animals; D000907:Antibodies, Bacterial / Q000276:immunology; D000942:Antigens, Bacterial / Q000235:genetics* / Q000378:metabolism; D001426:Bacterial Proteins / Q000235:genetics* / Q000378:metabolism; D003001:Cloning, Molecular; D058501:Enzyme-Linked Immunospot Assay; D005260:Female; D051379:Mice; D008807:Mice, Inbred BALB C; D009169:Mycobacterium tuberculosis / Q000235:genetics / Q000378:metabolism; D010957:Plasmids / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000235:genetics / Q000494:pharmacology; D019444:Vaccines, DNA / Q000276:immunology*
chemical_list: D000907:Antibodies, Bacterial; D000942:Antigens, Bacterial; D001426:Bacterial Proteins; C510537:MPT64 protein, Mycobacterium tuberculosis; D011993:Recombinant Fusion Proteins; D019444:Vaccines, DNA; D000217:Acyltransferases; C121433:antigen 85B, Mycobacterium tuberculosis