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

结核分枝杆菌ESAT-6和CFP-10抗原的克隆、表达和纯化

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作者:Shima Mahmoudi, Setareh Mamishi, Mona Ghazi, Reihaneh Hosseinpour Sadeghi, Babak Pourakbari

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.

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.

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