cloning, expression and purification of mycobacterium tuberculosis esat-6 and cfp-10 antigens.
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abstract
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.
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Journal title:
iranian journal of microbiologyجلد ۵، شماره ۴، صفحات ۳۷۴-۳۷۸
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