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

作　　者：石玉林 王博 姚蜜蜜 汪劲松[1] 

机构地区：[1]湖北师范学院生命科学学院,湖北黄石435002

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

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.

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 

分 类 号：Q51[生物学—生物化学]