重组人FS315C末端蛋白表达及其抗体制备  

The Expression of the Recombinant Human FS315 C Terminal Peptide and Preparation of Its Antibody

作　　者：杨煜[1] 劳凤学[1] 高振平[1] 徐桂月[1] 张鹏宇[1] 柳忠辉[1] 

机构地区：[1]吉林大学基础医学院吉林省神经免疫重点实验室,长春130021

出　　处：《中国生物工程杂志》2005年第8期56-59,共4页China Biotechnology

基　　金：吉林省科技厅资助项目(980580);吉林省杰出青年基金项目(4-20-5)

摘　　要：构建FS315C末端肽原核表达载体pGEX-FS315C,表达重组人卵泡抑素315(FS315)C末端肽,制备抗FS315C末端抗体。实验结果显示,重组pGEX-FS315C表达质粒在E.coli BL21中高表达GST-FS315C融合蛋白,采用亲和层析与电泳纯化GST-FS315C免疫家兔,获得抗FS315C末端抗体,Western blot检测显示该抗体只与FS315结合,而与FS288无交叉反应。ELISA检测显示抗FS315C末端抗体与FS315特异结合,而与FS288、inhibin及activin A等蛋白均无交叉反应。利用该抗体进行的免疫组化染色显示巨噬细胞FS表达阳性,与以往报道一致。实验采用重组GST-FS315C免疫家兔,成功地制备了抗FS315C末端特异抗体。To express the recombinant human FS315C terminal peptide and to prepare its antibody, the prokaryotic expression plasmid pGEX-FS315C for FS315C terminal genes was constructed, and the anti-human FS315 C terminal polyclonal antibody was prepared by immunizing New Zealand rabbit with FS315C protein. The results showed that the recombinant pGEX-FS315C plasmid highly expressed GST-FS315C fusion protein in E. coli BL21, and anti-FS315C terminal polyclonal antibody was obtained by immunizing New Zealand rabbit with GST-FS315C fusion protein purified by affinity chromatography and electrophoresis. The antibody against FS315C terminal only specifically binds to FS315 by Western blot, but has no binding with FS288. By ELISA, the antibody against FS315C terminal has specific reaction with FS315, but has no cross-reaction with FS288, inhibin or activin. The immunohistochemistry showed that FS was expressed in macrophages with the antibody to FS315C terminal, which was consistent with the previous report. The specific anti-FS315C terminal polyclonal antibody was successfully prepared by immunizing New Zealand rabbit with recombinant GST- FS315C.

关 键 词：FS315 GST融合蛋白 抗体 抗体制备 蛋白表达 重组人 免疫组化染色 Western 原核表达载体 E.COLI 

分 类 号：Q786[生物学—分子生物学] Q78