Cloning and expression of Mycobacterium tuberculosis ESAT-6 in prokaryotic system
2009
Tebianian, M., Tarbiat Modares University, Tehran, Iran | Zavaran Hoseini, A., Tarbiat Modares University, Tehran, Iran | Ebrahimi, S.M., Razi Vaccine and Serum Research Institute, Karaj, Iran | Rezaei Mokaram, A., Razi Vaccine and Serum Research Institute, Karaj, Iran | Taghizadeh, M., Razi Vaccine and Serum Research Institute, Karaj, Iran | Asli, E., Razi Vaccine and Serum Research Institute, Karaj, Iran
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 plasmid was constructed. This hybrid vector was transformed in E. coli M15 and expressed in optimal condition. The expressed protein was analyzed on SDS-PAGE and confirmed by western blotting using specific antisera to ESAT-6. We successfully cloned and expressed ESAT-6 (His)6 from M. tuberculosis H37Rv genome. As well as usage for serodiagnosis, this recombinant protein offers the potential development of other vaccine formats such as DNA or subunit vaccines against tuberculosis.
Показать больше [+] Меньше [-]