| 拟南芥VSP2蛋白的重组表达及其酸性磷酸酶活性的测定 |
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| 引用本文： | 王进,史竹兵,张敏.拟南芥VSP2蛋白的重组表达及其酸性磷酸酶活性的测定[J].生物学杂志,2009,26(6):53-55. |
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| 作者姓名： | 王进 史竹兵 张敏 |
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| 作者单位： | 安徽大学生命科学学院,合肥,230039 |
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| 摘 要： | 通过PCR扩增从拟南芥cDNA文库中得到VSP2蛋白的编码序列,将其构建到原核表达载体pET-22b上,并在大肠杆菌BL21菌株中实现高效可溶表达。经过Ni-NTA亲和层析一步纯化,获得电泳纯的重组VSP2蛋白。以pNPP为底物检测,该蛋白具有酸性磷酸酶活性,反应的最适pH值4.5,最适温度为45oC,Km值为26.2mM。重组VSP2蛋白表达量高,纯化后均一性好,适于蛋白晶体生长。
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| 关 键 词： | 拟南芥VSP2蛋白 克隆表达 酸性磷酸酶活性 晶体生长 |
Bacterial expression and acid phosphatase activity of VSP2 from Arabidopsis thaliana |
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| WANG Jin,SHI Zhu-bing,ZHANG Min.Bacterial expression and acid phosphatase activity of VSP2 from Arabidopsis thaliana[J].Journal of Biology,2009,26(6):53-55. |
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| Authors: | WANG Jin SHI Zhu-bing ZHANG Min |
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| Institution: | ( School of Life Science, Anhui University, Hefei 230039, China) |
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| Abstract: | The VSP2 cDNA sequence was amplified by PCR from cDNA library of Arabidopsis thaliana and cloned into pET-22b vector for bacterial expression. The soluble recombinant protein was over-expressed in E. coli BL21 and purified by a one-step Ni-NTA column chromatography method. The purified protein had acid phosphatase activity with general substrate pNPP. The optimum pH and temperature of reaction was 4.5 and 45℃, respectively. The Km was 26.2mM. The purified recombinant VSP2 had good homogeneity and was suitable for crystal growth. |
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| Keywords: | Arabidopsis thaliana VSP2 clone and expression acid phosphatase activity crystal growth |
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