Cloning, expression and purification of inhA from mycobacterium tuberculosis
- VernacularTitle:结核分枝杆菌inhA基因的克隆、表达及高纯度inhA蛋白的制备
- Author:
Shu CHEN
;
Wenhong ZHANG
;
Chaoneng JI
- Publication Type:Journal Article
- Keywords:
Mycobacterium tuberculosis;
Gene expression;
Purification
- From:
Chinese Journal of Infectious Diseases
2001;0(03):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective To clone and express inhA gene from mycobacterium tuberculosis , and purify the inhA protein. Methods Recombinant plasmid pET 24b/inhA was constructed and transferred into Escherichia coli . After restriction enzyme analysis and sequencing, the host bacteria were induced by IPTG and the product was identified by SDS PAGE. Furthermore, the overexpressed inhA protein was purified by Nit NTA Superflow system. Results The inhA gene was overexpressed in E. coli, the production was corresponding to 30 percent of total cell protein. Using Nit NTA Superflow,we can get more than 99% purified protein. Conclusions The cloning, expression and purification of inhA gene are successful.
