小鼠FGF9在大肠杆菌中的表达与纯化

为了建立小鼠FGF9的原核表达体系,获得纯化的FGF9重组蛋白,采用PCR技术扩增FGF9,并将其克隆入原核表达载体pET30a(+)中,经测序验证后将该重组质粒转入大肠杆菌BL21(DE3)中进行表达,采用亲和层析技术进行目的蛋白的纯化,并用Western blotting进行了免疫原性验证.测序结果显示插入到载体中的片段与FGF9的天然序列一致,在BL21(DE3)中表达出了可溶性的FGF9重组蛋白,其表达量达到总蛋白的40 %;用镍螯合亲和层析方法获得了纯度大于95 %的FGF9重组蛋白.

Expression and Purification of Mouse Fibroblast Growth Factor 9 in Eco.li

To express and purify mouse FGF9 peptide in Eco.li, the gene was amplified by PCR.,which was subsequently cloned into the plasmid pET30a(+).The insert was confirmed by DNA sequencing. The recombinant plasmid was transferred into BL21(DE3)to express recombinant FGF9 peptide. The recombinant peptid successfully purified by Nickel affinity chromatography,its immunological characteristics was analyzed by Western blotting technology. The sequnce analysis suggested the insert had same sequence coding for natural FGF9 protein. The recombinant protein inducible expressed by IPTG was soluble in Eco.li BL21 (DE3). The targeted protein expression rate accounted for about 40 % of total bacteria protein.The purity of target protein was above 95 % by using Nickel affinity chromatography.