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

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SspDnaB基因的质粒载体pTWIN1中，将SspdnaB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.