cloning, expression and purification of mycobacterium tuberculosis esat-6 and cfp-10 antigens.
نویسندگان
چکیده
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.
منابع مشابه
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 METH...
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Expression and Purification of Recombinant Mycobacterium Tuberculosis (TB) Antigens, ESAT-6, CFP-10 and ESAT- 6/CFP-10 and Their Diagnosis Potential for Detection of TB Patients
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The identification of a large number of antigens with potential for development of new tuberculosis vaccine has been accomplished in recent years. This study was designed for cloning and expression of ESAT-6 as a potent antigen of Mycobacterium tuberculosis. Selected gene (Rv3875) was amplified by PCR and product was ligated into expressing plasmid vector pQE30 and recombinant pQE30-ES plasmi...
متن کاملMacrophage Immune Response Suppression by Recombinant Mycobacterium tuberculosis Antigens, the ESAT-6, CFP-10, and ESAT-6/CFP-10 Fusion Proteins
BACKGROUND Macrophage immune responses are affected by the secretory proteins of Mycobacterium tuberculosis (Mtb). This study aimed to examine the immune responses of macrophages to Mtb secretory antigens, namely ESAT-6, CFP-10, and ESAT-6/CFP-10. METHODS THP-1 cells (a human monocytic cell line) were cultured and differentiated to macrophages by phorbol 12-myristate 13-acetate. The cytotoxic...
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عنوان ژورنال:
iranian journal of microbiologyجلد ۵، شماره ۴، صفحات ۳۷۴-۳۷۸
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