| 谷胱甘肽S-转移酶的诱导表达及提纯 |
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| 引用本文： | 杨丹,叶佩莹,万津,刘永平,马辉文.谷胱甘肽S-转移酶的诱导表达及提纯[J].氨基酸和生物资源,2004,26(3):33-37. |
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| 作者姓名： | 杨丹 叶佩莹 万津 刘永平 马辉文 |
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| 作者单位： | 1. 武汉大学药学院,湖北武汉,430072;2. 武汉大学生命科学学院,湖北武汉,430072 |
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| 摘 要： | 含有日本血吸虫谷胱甘肽S -转移酶基因的质粒 pGEX - 5X - 1在大肠杆菌BL2 1 (DE3)菌株中得到表达。 37℃1 ]下用 1 %乳糖诱导谷胱甘肽S -转移酶 (GST)的最适表达时间为 3h。盐析粗提酶液并以Habig法2 ] 监测GST的活性 ,用pH6 .0 ,饱和度为 30 %硫酸铵去除杂蛋白 ,并调 pH7.5 ,饱和度 70 %硫酸铵使GST大量析出并保持活性。用透析法脱盐 ,并用Sephadex -G5 0凝胶对GST进行了初步纯化。初步探讨了GST的保存方法
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| 关 键 词： | 谷胱甘肽S-转移酶 基因表达 GST的纯化 |
| 文章编号： | 1006-8376(2004)03-0033-05 |
| 修稿时间： | 2004年2月17日 |
Expression and Purification of Glutathione S-Transferase |
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| YANG Dan ,YE Pei-ying ,WAN Jin ,LIU Yong-ping ,MA Hui-wen.Expression and Purification of Glutathione S-Transferase[J].Amino Acids & Biotic Resources,2004,26(3):33-37. |
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| Authors: | YANG Dan YE Pei-ying WAN Jin LIU Yong-ping MA Hui-wen |
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| Institution: | YANG Dan 1,YE Pei-ying 2,WAN Jin 1,LIU Yong-ping 1,MA Hui-wen 1 |
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| Abstract: | 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. |
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| Keywords: | glutathione S-transferase gene expression purification of GST |
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