大鼠热休克因子结合蛋白1原核表达载体的构建及表达

Construction of pGEX-2TK-Hsbp1 Prokaryotic Expression Vector and Expression of HSBP1

以重组克隆载体pGEM—T—hsbp1为模板,PCR扩增hsbp1,用限制性内切酶双酶切、T4连接酶连接、转化BL -21的方法构建pGEX-2TK-hsbp1原核表达载体,实现了hsbp1在BL21中的表达,SDS—PAGE检测表达的融合蛋白主要以可溶形式存在,并利用亲和层析得到融合蛋白。

Using recombination cloning plasmid pGEM-T-hsbpl as template, heat shock factor binding protein gene was cloned ; By restriction endonuclease, T4 DNA ligase. BL-21, the pGEX-2TK-hsbpl prokaryotic expression vector was constructed. The recombinant expression vector can express fusion proteins of recombinant HSBP1 in BL21. The result of SDS-PAGE manifests that the target protein was expressed in soluble form. We obtained target protein by affinity chromatography.