华北农学报Issue(3)：31-36,6.DOI:10.7668/hbnxb.2015.03.006
绵羊 FecB 基因的原核表达及其抗体制备
Prokaryotic Expression of FecB Gene and Generation of Its Polyclonal Antibody
摘要
Abstract
Further study on FecB gene,the complete FecB gene was amplified with PCR method using a pair of specific primers designed according to the relevant nucleotide sequence from GenBank .And FecB gene was cloned into vector pET30a(+) for expressed in E.coli BL21(DE3).The expression of 6 ×His and FecB fusion protein was induced by IPTG ,then purified by Ni-NTA chromatographic method .Polyclonal antibodies were prepared by im-munized mice with the purified recombinant protein .Expression of the target protein was detected by SDS-PAGE, the specificity and titer of the antibody in anti-sera was determined by Western Blot and ELISA respectively .The re-combinant FecB can be expressed by IPTG induction and purified by Ni-NTA resin.The results of ELISA and West-ern Blot proved that polyclonal antiserum prepared with purified recombinant FecB as antigen has high titration (>1∶32 000 ) and specificity .A method for prokaryotic expression and purification of sheep FecB was established and the anti-FecB polyclonal antibody with high titration and specificity were obtained .These results would provide reli-able tools for the future study on FecB function .关键词
FecB基因/原核表达/蛋白纯化/多克隆抗体制备Key words
FecB gene/Prokaryotic expression/Protein purification/Polyclonal antibody分类
生物科学引用本文复制引用
丁强,张利平,贾建磊,王静,张勇..绵羊 FecB 基因的原核表达及其抗体制备[J].华北农学报,2015,(3):31-36,6.基金项目
绵羊双羔性状分子调控机理研究项目（ GNSW-2012～24）；通过蛋白表达差异与动态组成研究绵羊双羔性状的信号分子及其调控机理项目 （）
