小鼠激活素受体相互作用蛋白2的原核表达及抗血清制备

目的]克隆小鼠激活素受体相互作用蛋白2（ARIP2）cDNA序列，利用大肠杆菌表达ARIP2，制备兔抗小鼠ARIP2的多克隆抗血清。方法]采用RT-PER方法，将小鼠ARIP2基因克隆到pET-32a（＋）质粒，构建ARIP2原核表达载体并转化到大肠杆菌，IgrG诱导融合蛋白的表达，经亲和层析纯化获得可溶性重组蛋白，免疫家兔后制备ARIP2抗血清，分别采用ELISA，Western blot检测抗体效价和特异性。结果]测序结果表明重组质粒pET32a（＋）-ARIP2构建成功；SDS-PAGE分析表明获得的重组蛋白为可溶蛋白；经过亲和层析后得到有效分离；Elisa法测定的抗血清效价为1：50000；Westernblot鉴定结果表明抗血清能与目的蛋白特异性结合。结论]成功将ARIP2进行了原核表达并制备了高效价、高特异性的兔抗小鼠ARIP2抗血清，为进一步研究ARIP2功能提供了有力工具。

Prokaryotic Expression and Preparation of Antiserum for Mouse Activin Receptor-interacting Protein 2

Objective] The study aims to clone mouse activin receptor-interacting protein 2(ARIP2) sequences and express recombinant ARIP2 E.coil,and prepare high quality rabbit polyclonal antiserum.Method] The out-membrane fragment of ARIP2 gene is amplified by PCR from mouse liver cDNA pool,which is cloned into pET-32a(+) plasmid.E coli BL21(DE3) transformed with recombinant pET-ARIP2 plasmid is induced by IPTG to express ARIP2 in high level,expressed soluble ARIP2 is purified by NI-NTA agarose to immunize rabbit.T...