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《Amino Acids & Biotic Resources》 2004-03
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Expression and Purification of Glutathione S-Transferase

YANG Dan 1 , YE Pei-ying 2 ,WAN Jin 1 , LIU Yong-ping 1 ,MA Hui-wen 1 (1. College of pharmacy, Wuhan University,Wuhan430072,China; 2. College of life sciences,Wuhan University,Wuhan430072,China)  
Glutathione S-transferase (GST)gene of Schistosoma japonicum harboring in the plasmid pGEX-5X-1 was expressed in Escherichia coli BL21(DE3) induced with lactose. The optimal inducing time was 3 hours at the temperature of 37℃. The expressed GST was salted out by 70% saturated ammonium sulphate, dialysed and partially purified with Sephadex-G50 gel filtration-chromatography. The enzymatic activity of GST in the cell lysate was determined with spectrophotometericrely described by Habig et al. and the expressing level of GST evaluated with SDS-PAGE was used as indexes to optimize the process of induction and purification of GST.
【CateGory Index】: Q814
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