Cloning,expression and purification of Mycobacterium tuberculosis ESAT-6 and CFP-10 antigens

Background and objectives: esat-6 (6-kdaearly secretory antigenic target) and cfp-10 (10-kda culture filtrate protein) have been described as dominant antigens recognized by t-cells and considered as virulence factors in mycobacterium tuberculosis. the aim of this study was to clone,express and purify recombinant esat-6 andcfp-10 proteins of m. tuberculosis in soluble form. materials and methods: esat-6 andcfp-10 genes were amplified by pcr,cloned into pet32a (+) vector,and overexpress-ed using isopropyl-beta-d-thiogalactopyranoside in e. coli bl21 (de3). esat-6 andcfp-10 proteins were purified by ni-nta affinity chromatography and were detected by anti- esat-6 and anti -cfp10 antibodies. results: esat-6 andcfp-10 genes were successfully expressed and purified. anti- esat-6 and anti-cfp-10 antibodies were produced after induction of immunization against purified esat-6 andcfp-10 proteins in rabbit. conclusion: in this study,we cloned,expressed and purified sufficient amounts of esat-6 andcfp-10 and it would be tested for the development of diagnostic kit for m. tuberculosis in future.