猪抵抗素(resistin)的原核表达及表达产物的纯化鉴定

用PCR方法扩增到抵抗素基因(Resistin, RSTN)并将其亚克隆至pET-32a(+)表达载体，获得重组质粒pET-RSTN。将重组质粒转化大肠杆菌BL-21（DE3）感受态细胞，用IPTG诱导表达。SDS-PAGE检测结果表明，重组resistin蛋白分子量大小约30kDa。对表达条件如温度、IPTG浓度及诱导时间进行优化并用SDS-PAGE检测。结果表明，30℃、4h、IPTG浓度为1mmol/L时，可溶性重组resistin的含量最高。表达产物经Western blot检测证实是Resistin蛋白，并用镍离子亲和层析的方法获得纯化的Resistin蛋白。

The prokaryotic expression of Porcine Resistin and its purification, identification

Resistin is a newly discovered adipocyte hormone.With the method of gene engineering,we got resistin.The RSTN gene was subcloned to the expression vector pET-32a(+) and the recombined plasmid named pET-RSTN was constructed successfully. The plasmid pET-RSTN were transformed into the Eoli.BL-21(DE3). After inducing by IPTG the fusion protein of resistin were expressed and its molecular weight was about 30kDa. As temperature、IPTG concentration and induction time were optimized,the soluble recombinant protein was high expressed at 30℃ for 4h with 1mmol/L IPTG induction. The product was identified by SDS-PAGE and western-blotting. The expression production purified on Ni2+-NTA column.