Cloning, expression and purification of human glutathione S-transferase Pi
WANG Zhihui 1,2) , WU Yiming 1) ,WU Yongjun 1) 1)Department of Occupational Health, College of Public Health, Zhengzhou University, Zhengzhou 450052 2)Department of Occupational Health,Henan Institution of Professional Medicine, Zhengzhou 450052
Aim: To express and purify the human Pi class glutathione S-transferase (hGST Pi). Methods: The expression of plasmid for hGST Pi was constructed by ligation of the cDNA that codes for the protein into the expression vector pMAL-C2x.The expressed protein was purified by maltose affinity column chromatography, which produced the pure and active enzyme in one step. Results: The activity of the purified protein was 45 mmol/(min·mg protein) from the maltose affinity column. The purity of the protein was assessed by maltose-tagged chromatography. It showed that the real molecular weight of the maltose-tagged hGST Pi polypeptide chain agreed with the calculated value and that the purified protein eluted as anapparent homodimer on the gel filtration column. Conclusion: Our expression system allows the expression and purification of active maltose-tagged hGST Pi in high yield with no need of removal of the maltose tag and gives pure protein in one purification step towards further study of this enzyme.
【CateGory Index】： R346