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

被引：0
|
作者：
Mahmoudi, Shima [1 ]
Mamishi, Setareh [1 ,2 ]
Ghazi, Mona [3 ]
Sadeghi, Reihaneh Hosseinpour [1 ]
Pourakbari, Babak [1 ]
机构：
[1] Univ Tehran Med Sci, Pediat Infect Dis Res Ctr, Tehran, Iran
[2] Univ Tehran Med Sci, Children Med Ctr Hosp, Sch Med, Dept Infect Dis, Tehran, Iran
[3] Shahid Beheshti Univ Med Sci, Sch Med, Dept Microbiol, Tehran, Iran
来源：
IRANIAN JOURNAL OF MICROBIOLOGY | 2013年 / 5卷 / 04期
关键词：
M; tuberculosis; ESAT-6; CFP-10;
D O I：
暂无
中图分类号：
Q93 [微生物学];
学科分类号：
071005 ; 100705 ;
摘要：
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 and CFP-10 proteins of M. tuberculosis in soluble form. Materials and Methods: ESAT-6 and CFP-10 genes were amplified by PCR, cloned into pET32a (+) vector, and overexpressed 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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页码：374 / 378
页数：5
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