莱茵衣藻缺铁应答调控基因femu2p-C2H2的融合表达与纯化

Fusion Expression and Purification of Femu2p-C2H2 Gene Regulating and Controlling Iron Deficiency Response in Chlamydomonas reinhardtii

为获得目标蛋白以用于蛋白功能解析,本研究将锌指蛋白C2H2以GST(Glutathione S-transferase)融合蛋白的形式进行了表达,并确定了表达的最佳条件:37℃、IPTG诱导4 h、IPTG终浓度为1.0 mmol/L。利用固定化的还原型谷胱甘肽(GSH)对GST-C2H2融合蛋白进行一步亲和纯化,获得目标融合蛋白,25倍超滤浓缩之后测定其浓度为1.762 mg/mL。

In order to obtain target protein for protein fimction analysis, this study expressed the zinc finger protein C2H2 with GST （Glutathione S-transferase） fusion protein and determined the best condition of the fusion protein expression： 37℃, induced by IPTG for 4 hours and the IPTG final concentration was 1.0 mM. Using immobilized and reduced glutathione （GSH） purified the GST-C2H2 fusion protein, target fusion protein could be obtained. The concentration of the GST-C2H2 fusion protein is 1.762 mg/ml after 25 times concentrated by ultrafiltration.