类法尼醇X受体α基因的原核表达及多克隆抗体制备  

Prokaryotic expression and antibody preparation of farnesoid X receptor α

作　　者：李超燕[1] 于琨[1] 朱海华 张玉娟[1] 顾伟祯 郑文明[1] 许君[1] 

机构地区：[1]河南农业大学生命科学学院,河南郑州450002 [2]河南省商业科学研究所有限责任公司,河南郑州450002

出　　处：《河南农业大学学报》2015年第2期244-248,共5页Journal of Henan Agricultural University

基　　金：国家自然科学基金资助项目(31201332);河南省科技攻关重点项目(132102310033)

摘　　要：根据NCBI上公布的序列设计引物,从Hep G2细胞系中克隆了FXRα的开放阅读框(ORF),构建原核表达载体p ET-28a-FXRα,转化E.coli BL21(DE3),经1.0 mmol·L-1IPTG诱导表达,SDS-PAGE检测,成功诱导FXRα蛋白;以镍柱亲和层析纯化获得FXRα蛋白,免疫家兔,制备多克隆抗体。Western Blot分析表明:制备的多克隆抗体能够特异识别重组蛋白,为深入研究此受体的功能及新药发现奠定了基础。The open reading frame（ ORF） sequence for FXRα was amplified by RT-PCR form Hep G2 cell line. The sequence was sub-cloned into vector p ET-28 a for expression in E. coli BL21（ DE3）.The expression of recombinant fusion FXRα with 6 × His-tag was induced by isopropylthio-β-D-galactoside（ IPTG）. The expression of the target protein was detected by SDS-PAGE and then purified by Ni NTA chelating agarose. SDS-PAGE analysis showed that FXRα protein was success fully expressed in E. coli BL21（ DE3）. A rabbit polyclonal antibody against recombinant FXRα was obtained by immunization rabbit with the purified FXRα protein. It was showed that the anti-sera could bind to the expressed protein specifically by Western blot analysis. This work provides a foundation for further study on receptor function and new drug development.

关 键 词：类法尼醇X受体α 核受体 原核表达 抗体制备 

分 类 号：Q78[生物学—分子生物学]