| 尿激酶催化结构域在毕氏酵母中的表达、纯化及结晶 |
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| 引用本文： | 赵更香,袁 彩,卞传兵,江龙光,叶晓明,黄子详,黄明东.尿激酶催化结构域在毕氏酵母中的表达、纯化及结晶[J].生物化学与生物物理进展,2006,33(4):377-381. |
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| 作者姓名： | 赵更香 袁 彩 卞传兵 江龙光 叶晓明 黄子详 黄明东 |
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| 作者单位： | 中国科学院福建物质结构所,结构化学国家重点实验室,中国科学院研究生院,福州,350002 |
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| 基金项目： | 国家高技术研究发展计划(863计划);国家重点实验室基金;中国科学院"百人计划" |
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| 摘 要： | 利用重叠延伸PCR方法扩增尿激酶催化结构域的突变体基因片段,将其克隆至表达载体pPICZαA上,转化酵母X-33,用Zeocin筛选高拷贝数的酵母菌落.重组蛋白通过阳离子琼脂糖柱纯化,纯度达到99%,该仅含尿激酶催化结构域的突变体(C279A/N302Q),无需激活即具有尿激酶活性.用气相扩散法获得蛋白质晶体,其衍射分辨率达1.45!.
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| 关 键 词： | 尿激酶 Pichiapastoris 纤溶酶原 重叠延伸PCR 结晶 |
| 收稿时间： | 2005-11-08 |
| 修稿时间： | 2005-11-082005-12-31 |
Expression, Purification and Crystallization of Urokinase Catalytic Domain in Pichia pastoris |
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| ZHAO Geng-Xiang,YUAN Cai,BIAN Chuan-Bing,JIANG Long-Guang,YE Xiao-Ming,HUANG Zi-Xiang and HUANG Ming-Dong.Expression, Purification and Crystallization of Urokinase Catalytic Domain in Pichia pastoris[J].Progress In Biochemistry and Biophysics,2006,33(4):377-381. |
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| Authors: | ZHAO Geng-Xiang YUAN Cai BIAN Chuan-Bing JIANG Long-Guang YE Xiao-Ming HUANG Zi-Xiang and HUANG Ming-Dong |
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| Institution: | State Key Laboratory of Structural Chemistry, Graduate University of The Chinese Academy of Sciences, Fujian Institute of Research on The Structure of Matter, The Chinese Academy of Sciences, Fuzhou 350002, China;State Key Laboratory of Structural Chemistry, Graduate University of The Chinese Academy of Sciences, Fujian Institute of Research on The Structure of Matter, The Chinese Academy of Sciences, Fuzhou 350002, China;State Key Laboratory of Structural Chemistry, Graduate University of The Chinese Academy of Sciences, Fujian Institute of Research on The Structure of Matter, The Chinese Academy of Sciences, Fuzhou 350002, China;State Key Laboratory of Structural Chemistry, Graduate University of The Chinese Academy of Sciences, Fujian Institute of Research on The Structure of Matter, The Chinese Academy of Sciences, Fuzhou 350002, China;State Key Laboratory of Structural Chemistry, Graduate University of The Chinese Academy of Sciences, Fujian Institute of Research on The Structure of Matter, The Chinese Academy of Sciences, Fuzhou 350002, China;State Key Laboratory of Structural Chemistry, Graduate University of The Chinese Academy of Sciences, Fujian Institute of Research on The Structure of Matter, The Chinese Academy of Sciences, Fuzhou 350002, China;State Key Laboratory of Structural Chemistry, Graduate University of The Chinese Academy of Sciences, Fujian Institute of Research on The Structure of Matter, The Chinese Academy of Sciences, Fuzhou 350002, China |
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| Abstract: | |
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| Keywords: | urokinase Pichia pastoris plasminogen site-mutated PCR crystallization |
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