Mycobacterium tuberculosis katG protein expression and purification of high.doc

 PAGE \* MERGEFORMAT 12 Mycobacterium tuberculosis katG protein expression and purification of high Keywords:: Mycobacterium tuberculosis; katG; gene expression; protein purification [Abstract] Objective Expression and purification of Mycobacterium tuberculosis katG protein, a mechanism for in-depth study of isoniazid-resistant foundation. Methods containing katG gene in pET24b-katG expression vector transformed into E. coli BL21 (DE3) strain, in the isopropyl-β -D-thiogalactoside (IPTG) induced expression, respectively, the expression of different induction time product was SDSand Coomassie brilliant blue staining. To obtain a stable high expression strain to be used after Xpress TM protein purification system to purify the ultrasonic breaking bacilli. Finally, the product was purified catalase activity of the initial testing. Results after induction of recombinant E. coli bacilli to sodium dodecyl sulfate - polyacrylamide gel electrophoresis (SDS) and Coomassie brilliant blue staining revealed that the relative molecular mass of approximately 80 000. The amount of expressed protein amount of about 17.7% of total protein. KatG gene recombinant product was purified and found to 350 mmol / L imidazole elute the best in the purification, protein purity above 90%. Expressed product of the catalase activity of the initial testing proved that the reorganization of katG gene product with hydrogen peroxide activity. [HTH〗 Conclusion pET24b-katG expression plasmid was transformed into E. coli katG available in high expression of recombinant strains, the expression products have a certain activity, after purification can be achieved through a higher purity. Overexpression and purification of catalase-peroxidase katG from mycobacterium tuberculosis [Abstract] Objective To express and purify the catala se-peroxidase katG gene from mycobacterium tuberculosis. Methods Plasmid pET24b containing katG was transferred into com petent Escherichia coli and katG gene was