Tehran University of Medical Sciences Iranian Journal of Microbiology 2008-3289 5 4 2013 12 15 374 378 EN Shima Mahmoudi Pediatric Infectious Disease Research Center, Tehran University of Medical Sciences, Tehran, Iran. Setareh Mamishi Pediatric Infectious Disease Research Center, Tehran University of Medical Sciences, Tehran, Iran AND Department of Infectious Disease, Children Medical Center Hospital, School of Medicine, Tehran University of Medical Sciences, Tehran, Iran. Mona Ghazi Department of Microbiology, School of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran. Reihaneh Hosseinpour Sadeghi Pediatric Infectious Disease Research Center, Tehran University of Medical Sciences, Tehran, Iran. Babak Pourakbari Pediatric Infectious Disease Research Center, Tehran University of Medical Sciences, Tehran, Iran. 2015 10 16 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. https://ijm.tums.ac.ir/index.php/ijm/article/view/501 https://ijm.tums.ac.ir/index.php/ijm/article/download/501/276