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、位点特异性蛋白酶切得到纯化的Atox1蛋白 The cDNA fragment of human transfer copper chaperone Atox1 was amplified by PCR and directly cloned into PCRII vector. After DNA sequencing, it was inserted into GST fusion expression vector PGEX 6p 2, The recombinant plasmid pGEA was constructed. The plasmid was introduced into Escherichia coli. The fusion protein of PGEA fusion protein induced by IPTG was purified by affinity chromatography. The purified Atox1 protein was obtained by site-specific protease digestion