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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

The expression and purification of the N-domain of arginine kinase mediated by the intein SspdnaB
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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。 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.
出处 《湖北师范学院学报(自然科学版)》 2011年第4期86-91,共6页 Journal of Hubei Normal University(Natural Science)
基金 湖北省自然科学基金(2005AB192) 黄石市2010年度科技计划项目资助
关键词 精氨酸激酶 蛋白质内含子却DnaB N端结构域 融合蛋白 蛋白质剪切 镍亲和层析 arginine kinase N- domain intein Ssp DnaB fusion protein protein splicing Ni affinity chromatography
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