人体铜伴侣蛋白Atox1在大肠杆菌中的表达

Expression of human Copper Chaperone Protein Atox1 in E.coli

利用PCR技术扩增出人体转铜伴侣蛋白Atox1的cDNA片段 ,直接克隆到PCRII载体上 ,经DNA序列测定后 ,再插入到谷胱甘肽巯基转移酶 (GST)融合表达载体PGEX 6p 2上 ,构成重组表达质粒PGEA ,将此质粒导入大肠杆菌 ,经IPTG诱导后获得PGEA融合蛋白的表达 表达的融合蛋白经亲和层析。

cDNA of human copper chaperone Atox1 was amplified by PCR and cloned into PCRII vector After DNA sequencing, it was cloned into GST fusion protein expressive vector PGEX 6p 2. The recombinant plasmid PGEA was constructed and introduced into E coli GST Atox1 fusion protein was expressed at the induction of IPTG and was purified by affinity chromatography By cleavage of site specific protease and second affinity chromatography, GST was removed and recombinant Atox1 protein was purified