GST-Dot4融合蛋白的表达·纯化及鉴定

目的]为了提高Dotg蛋白在大肠杆菌中的可溶性表达并获得较高纯度的蛋白。方法]以pGEX—KG—Dot4重组质粒转化EcoliBL21后，用IPTG诱导重组菌表达GST—Dot4融合蛋白，采用SDS—PAGE及WesternBlot法鉴定表达产物，用GlutathioneSepharose4B亲和层析柱分离纯化。结果]pGEX—KG—Dot4重组表达载体在大肠杆菌中获得高效表达；经GlutathioneSepharose4B亲和层析柱纯化获得了高纯度的Dot4融合蛋白；WesternBlot表明，该蛋白可与GST标签抗体反应，表明获得了目的蛋白。结论]在大肠杆菌中表达纯化后得到较高纯度的GST—Dot4融合蛋白，为Dot4蛋白的结构、功能及作用机制研究奠定了基础。

Expression,Purification and Identification of GST-Dot4 Fusion Protein

Objective] The paper aimed to improve the soluble expression of Dot4p in E.coli and to obtain higher purity protein.Method]The pGEX-KG-Dot4 recombinant plasmid was transformed to E.coli BL2l and the expression of GST-Dot4 fusion protein was induced with IPTG.The expression product was identified by SDS-PAGE and Western Blot,and separated and purified by Glutathione Sepharose4B affinity chromatography column.Result] The pGEX-KG-Dot4 recombinant expression vector was successfully expressed in E.coli;and ob...