人FBXO31蛋白的原核表达及抗体制备  

Prokaryotic Expression of Human FBXO31 and Preparation of its Polyclonal Antibody

作　　者：黄海丽[1] 郑文岭[1] 赵蕊[1] 严启涛[1] 蔡翠霞[1] 张宝[1] 马文丽[1] 

机构地区：[1]南方医科大学基因工程研究所,广州510515

出　　处：《中国生物工程杂志》2010年第7期24-27,共4页China Biotechnology

摘　　要：FBXO31位于人染色人体16q24.3,是一个乳腺癌候选抑癌基因。为了获得抗FBXO31的多克隆抗体,构建了FBXO31基因的原核表达质粒pET-28a-FBXO31。将原核表达质粒pET-28a-FBXO31转入大肠杆菌BL21(DE3)菌株中,IPTG诱导表达融合蛋白。融合蛋白主要以包涵体形式存在,常规方法纯化包涵体。将纯化的pET-28a-FBXO31融合蛋白用于免疫新西兰大白兔获得抗血清,用pET-28a-FBXO31对抗血清进行分离纯化,得到抗FBXO31多克隆抗体。用Western印迹和免疫沉淀对其进行初步鉴定。获得了兔抗人FBXO31的多克隆抗体,为进一步研究FBXO31的生物功能提供了有用的工具。FBXO31 is the chromosome 16q24.3 senescence gene, a candidate breast tumor suppressor, and a component of an SCF complex. In order to obtain rabbit anti-FBXO31 polyclonal antibody, the complete coding sequence was cloned into pET-28a. Prokaryotic expression plasmid pET-28a-FBXO31 was constructed and expressed in the strain of E. coli BL21（DE3）. Fusion protein mainly existed in inclusion body. Fusion protein was purified and immune rabbit to obtain rabbit anti-FBXO31 polyclonal antibody. The specificity confirmed by Western blot and immunoprecipitation. The results showed that rabbit anti-FBXO31 polyclonal antibody could identify FBXO31 protein and be a useful tool in FBXO31 function analysis.

关 键 词：FBX031 原核表达 多克隆抗体 

分 类 号：Q786[生物学—分子生物学]