人源核受体hLRH-1 的表达纯化及多克隆抗体制备

目的：制备抗人源核受体hLRH-1 的多克隆抗体，为进一步研究其功能奠定基础。方法：构建含有hLRH-1基因全长克隆的 原核表达载体pET507a-hLRH-1 并用IPTG 诱导其在Rosseta2 菌株中表达重组蛋白His-hLRH-1，经亲和层析纯化后按常规方法 免疫新西兰兔制备多克隆抗体，并用Western Blot 对其特异性进行鉴定。结果：原核表达载体pET507a-hLRH-1经测序证实构建 成功，将其转化大肠杆菌Rosseta2菌株后成功诱导表达重组蛋白His-hLRH-1，经纯化免疫新西兰兔后得到抗hLRH-1 多克隆抗 体，Western blot 证实抗体具有高度特异性。结论：成功表达His-hLRH-1 重组蛋白并制备出多克隆抗体，为进一步用于hLRH-1 的 免疫学检测及其功能研究奠定了基础。

Expression, Purification and Polyclonal Antibody Preparation of Human Nuclear Receptor hLRH-1

Objective:To generate rabbit polyclonal antibody against human nuclear receptor hLRH-1, which could facilitate its functional study.Methods:The recombinant plasmid pET507a-hLRH-1 is constructed and transformed into Rosseta2, the expression of His-hLRH-1 recombinant protein is induced by IPTG. After purification by affinity chromatography, the recombinant protein is injected into New Zealand rabbits to generate polyclonal antibody against hLRH-1, the specificity is identified by Western Blot.Results:pET507a-hLRH-1 is constructed and the recombinant protein His-hLRH-1 is successfully expressed in Rosseta2. The anti- hLRH-1 rabbit polyclonal antibody is obtained and its high specificity is indicated by Western Blot.Conclusion:Rabbit polyclonal antibody against hLRH-1 is successfully produced, it will be a useful tool in future functional investigation of hLRH-1.