title: Cloning, expression and purification of Mycobacterium tuberculosis ESAT-6 and CFP-10 antigens.
abstract: OBJECTIVE
  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.
  
  
  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 -CFP-10 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.
  
  
  CONCLUSIONS
  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.
authors.affiliation: Pediatric Infectious Disease Research Center, Tehran University of Medical Sciences, Tehran, Iran.
authors.forename: Shima
authors.initials: S
authors.lastname: Mahmoudi
authors.affiliation: Pediatric Infectious Disease Research Center, Tehran University of Medical Sciences, Tehran, Iran ; Department of Infectious Disease, Children Medical Center Hospital, School of Medicine, Tehran University of Medical Sciences, Tehran, Iran.
authors.forename: Setareh
authors.initials: S
authors.lastname: Mamishi
authors.affiliation: Department of Microbiology, School of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran.
authors.forename: Mona
authors.initials: M
authors.lastname: Ghazi
authors.affiliation: Pediatric Infectious Disease Research Center, Tehran University of Medical Sciences, Tehran, Iran.
authors.forename: Reihaneh
authors.initials: R
authors.lastname: Hosseinpour Sadeghi
authors.affiliation: Pediatric Infectious Disease Research Center, Tehran University of Medical Sciences, Tehran, Iran.
authors.forename: Babak
authors.initials: B
authors.lastname: Pourakbari
keywords: CFP-10; ESAT-6; M. tuberculosis
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