| 新型载体蛋白P64K在大肠杆菌中的可溶性表达与纯化 |
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| 引用本文： | 熊向华,赵洪亮,薛冲,张伟,杨秉芬,刘志敏.新型载体蛋白P64K在大肠杆菌中的可溶性表达与纯化[J].生物技术通讯,2004,15(3):231-234. |
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| 作者姓名： | 熊向华 赵洪亮 薛冲 张伟 杨秉芬 刘志敏 |
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| 作者单位： | 军事医学科学院,生物工程研究所,北京,100071 |
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| 基金项目： | 国家重大科技专项(2002AA2Z345B) |
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| 摘 要： | 用常规PCR方法从脑膜炎球菌中克隆出P64K基因，构建重组质粒pET28a-P64K转化BL21(DE3)宿主菌，经眦诱导表达筛选P64K蛋白的高表达菌株。结果证实P64K蛋白为胞内可溶性表达，表达量约占胞内总蛋白的30％。重组P64K蛋白经Butyl疏水柱、G200分子筛柱和Q阴离子柱三步层析后纯度达到95％以上，为今后进一步的功能和应用研究打下了良好的基础。
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| 关 键 词： | 载体蛋白 P64K 大肠杆菌 可溶性表达 纯化 重组质粒 |
| 文章编号： | 1009-0002(2004)03-0231-04 |
| 修稿时间： | 2004年1月5日 |
The soluble expression in E.coli and purification of a new carrier protein, P64K |
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| XIONG Xiang-hua,ZHAO Hong-liang,XUE Chong,ZHANG Wei,YANG Bing-fen,LIU Zhi-min Institute of Biotechnology,Academy of Military Medical Sciences,Beijing,China.The soluble expression in E.coli and purification of a new carrier protein, P64K[J].Letters in Biotechnology,2004,15(3):231-234. |
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| Authors: | XIONG Xiang-hua ZHAO Hong-liang XUE Chong ZHANG Wei YANG Bing-fen LIU Zhi-min Institute of Biotechnology Academy of Military Medical Sciences Beijing China |
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| Institution: | XIONG Xiang-hua,ZHAO Hong-liang,XUE Chong,ZHANG Wei,YANG Bing-fen,LIU Zhi-min Institute of Biotechnology,Academy of Military Medical Sciences,Beijing100071,China |
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| Abstract: | P64K gene was amplified from Neisseria meningitids by using PCR and fused to the plasmid pET28a,then the recombinant plasmid was transformed into host cell BL21(DE3)and the high expressive clone was screened by using ex-pression test of IPTG induction.P64K was expressed as high level as a soluble form,accounting for approximately30%of the total cell protein.After three step chromatograhpy comprising of butyl hydrophobic,G200gel filtration and Q anion ex-change,the P64K protein was purified to appromixately95%. |
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| Keywords: | carrier protein P64K soluble expression |
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