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人FGF21原核表达载体的构建及重组蛋白表达

DOI: 10.13560/j.cnki.biotech.bull.1985.2015.04.032, PP. 218-222

Keywords: 人成纤维细胞生长因子(FGF21),克隆,原核表达,蛋白纯化

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Abstract:

构建人FGF21(fibroblastgrowthfactor,FGF)cDNA的原核表达载体并诱导其重组蛋白表达。提取人肝脏总RNA后,经RT-PCR扩增获得目的片段,构建其T载体进行保存。再构建重组原核表达载体pET-28a(+)-hFGF21,重组质粒转化至大肠杆菌菌株BL21(DE3)中,在IPTG诱导下得到可溶性表达,采用亲和层析法纯化表达产物后,进行Westernblot鉴定。成功构建重组质粒pET-28(+)-hFGF21,对其进行可溶性表达后成功纯化出his-hFGF21,经Westernblot鉴定该融合蛋白可与FGF21抗体特异性结合。成功构建pET-28(+)-hFGF21,并可溶性表达his-hFGF21蛋白。

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