| Ssp dnaB蛋白质内含子介导的精氨酸激酶N末端结构域的表达 |
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| 引用本文： | 石玉林,王博,姚蜜蜜,汪劲松.Ssp dnaB蛋白质内含子介导的精氨酸激酶N末端结构域的表达[J].湖北师范学院学报(自然科学版),2011,31(4):86-91. |
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| 作者姓名： | 石玉林 王博 姚蜜蜜 汪劲松 |
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| 作者单位： | 湖北师范学院生命科学学院 |
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| 基金项目： | 湖北省自然科学基金，黄石市2010年度科技计划项目资助 |
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| 摘 要： | 采用聚合酶链式反应(PCR)扩增精氨酸激酶(Arginine kinase,AK)N端基因片段,经双酶切后连接至含有蛋白质内含子Ssp DnaB基因的质粒载体pTWIN1中,将Ssp dnaB和N-domain的融合基因利用双酶切手段重组到含有组氨酸标签(His-tag)的质粒载体pET-28a中,得到含有组氨酸标签的融合蛋白基因,并在大肠杆菌Rosetta中表达,为获得具有天然氨基酸序列的精氨酸激酶N端结构域打下基础。
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| 关 键 词： | 精氨酸激酶 蛋白质内含子Ssp DnaB N端结构域 融合蛋白 蛋白质剪切 镍亲和层析 |
The expression and purification of the N- domain of arginine kinase mediated by the intein Ssp dnaB |
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| SHI Yu-lin,WANG Bo,YAO Mi-mi,WANG Jin-song.The expression and purification of the N- domain of arginine kinase mediated by the intein Ssp dnaB[J].Journal of Hubei Normal University(Natural Science),2011,31(4):86-91. |
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| Authors: | SHI Yu-lin WANG Bo YAO Mi-mi WANG Jin-song |
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| Institution: | (College of Life Science,Hubei Normal University,Huangshi 435002,China) |
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| Abstract: | AK N -domain＇s gene sequence has been amplified by PCR and connected to the pTWIN1 plasmid, which contains the gene of ssp dnaB. Then the fusion gene has been connected to the pET -28a plasmid in the same way. After the recombinant plasmid has been built, the fusion protein could be obtained by the Ni affinity chromatography, which provides us a sound preparation for getting nature AK N - domain efficiently. |
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| Keywords: | arginine kinase N- domain intein Ssp DnaB fusion protein protein splicing Ni affinity chromatography |
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