Rheb1基因多克隆抗体的制备

Preparation of Rheb1 Polyclonal Antibody

目的:在大肠杆菌BL21中表达并纯化GST-Rheb1融合蛋白,以之免疫成年新西兰大白兔,获得Rheb1多克隆抗体。方法:运用PCR,限制性内切酶酶切及T4连接酶连接的方法,构建带有Rheb1基因的真核和原核表达载体;IPTG诱导GST融合蛋白高表达,通过皮下注射免疫新西兰大白兔,并取血清进行免疫印记分析抗体效价。结果:成功的制备了高效价的Rheb1兔多克隆抗体。结论:Rheb1多克隆抗体的成功制备,将为研究Rheb1基因的功能发挥重要的作用。

Objective：To express and purify GST-Rhe1 fusion protein in E.coli BL21,and immunize the new zealand white rabbits with the fusion protein to get the polyclonal antibody.Methods：To make prokaryotic and eukaryotic expression vectors with PCR,restriction enzymes and T4 ligase.Purify the GST fusion protein induced by IPTG,and immunize the new zealand white rabbits with the fusion protein by subcutaneous injection.Identify the titer of the antibody by western bolt with mouse brain extract.Results：Succeed in preparation of high titer Rheb1 polyclonal antibody.Conclusion：The availability of recombinant protein and specific polyclonal antibodies will provide a valuable tool in studying biological function fo Rheb1.