| Expression and Purification of ZNF191（243-368） in Three Expression Systems |
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| 引用本文： | 赵东欣,滕欣辰,丁志春,黄仲贤.Expression and Purification of ZNF191（243-368） in Three Expression Systems[J].中国化学,2007,25(11):1739-1742. |
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| 作者姓名： | 赵东欣 滕欣辰 丁志春 黄仲贤 |
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| 作者单位： | [1]Department of Chemistry, Fudan University, Shanghai 200433, China [2]School of Chemistry and Chemical Engineering, Henan University of Technology, Zhengzhou, Henan 450001, China |
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| 基金项目： | Project supported by the National Natural Science Foundation of China （Nos. 30170228, 39990600）.Acknowledgements We thank Prof. Long Yu （Fudan University） for the ZNF191 gene. |
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| 摘 要： | ZNF191 （243-368）, a new human zinc finger protein, probably relates to some hereditary diseases and cancers, To obtain adequate amount of ZNF191（243-368） for the study of its property, structure and function, three different expression systems of inclusion-body, glutathione S-transferase （GST）, and hexahistidine （6 × His） were used and compared. Among these systems, the expression level of ZNFI91（243-368） was increased in inclusion body system under a higher isopropylthio-β-D-galactoside （IPTG） concentration, but the non-target proteins were also increased more, which made its purification more difficult and the yield lower. The expression of His-tag fusion protein was almost not affected by IPTG concentration, temperature and inducing time. At a high IPTG concentration the highest expression yield for GST fusion protein was obtained. And the fusion proteins can be partially purified by a single affinity chromatography step. The fusion protein systems show advantages for expression of these proteins.
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| 关 键 词： | 蛋白质表达 亲合色谱法 融合蛋白 半乳糖水解 |
| 修稿时间： | 2006-09-20 |
Expression and Purification of ZNF191(243–368) in Three Expression Systems |
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| ZHAO, Dong-Xin TENG, Xin-Cheng DING, Zhi-Chun HUANG, Zhong-Xian.Expression and Purification of ZNF191(243–368) in Three Expression Systems[J].Chinese Journal of Chemistry,2007,25(11):1739-1742. |
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| Authors: | ZHAO Dong-Xin TENG Xin-Cheng DING Zhi-Chun HUANG Zhong-Xian |
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| Institution: | 1 Department of Chemistry, Fudan University, Shanghai 200433, China ;2 School of Chemistry and Chemical Engineering, Henan University of Technology, Zhengzhou, Henan 450001, China |
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| Abstract: | ZNF191(243–368), a new human zinc finger protein, probably relates to some hereditary diseases and cancers. To obtain adequate amount of ZNF191(243–368) for the study of its property, structure and function, three different expression systems of inclusion-body, glutathione S-transferase (GST), and hexahistidine (6×His) were used and compared. Among these systems, the expression level of ZNF191(243–368) was increased in inclusion body system under a higher isopropylthio-β-D-galactoside (IPTG) concentration, but the non-target proteins were also increased more, which made its purification more difficult and the yield lower. The expression of His-tag fusion protein was almost not affected by IPTG concentration, temperature and inducing time. At a high IPTG concentration the highest expression yield for GST fusion protein was obtained. And the fusion proteins can be partially purified by a single affinity chromatography step. The fusion protein systems show advantages for expression of these proteins. |
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| Keywords: | protein expression affinity chromatography fusion protein isopropylthio-β-D-galactoside |
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