中国畜牧兽医2011，Vol.38Issue(8)：131-134,4.
奶牛黏附分子MadCam-1基因的克隆及原核表达
Cloning and Prokaryotic Expression of MadCam-1 Gene from the Dairy Cows
摘要
Abstract
To construct expression vertor of MadCam-1 gene, to expression and purify recombination protein and to prepare polyclonal antibody, which have laid a foundation of studying its function. A pair of primers was designed according to the sequence of MadCam-1 from GenBank. The MadCam-1 gene was amplified by PCR. The expression vector pGEX-4T-l-Mad-Cam-1 was reconstructed and transformed into E. coli BL21 ,then it was induced by IPTG. The expressed product was identified by SDS-PAGE and Western blotting. Rabbits were immunized with the purified MadCam-1 to generate antisera. The results showed that the homology of 100% of the cloned MadCam-1 gene to that reported in GenBank. The recombinant plasmid pGEX-4T-1-MadCam-1 was constructed successfully. The expression product was identified by Western blotting. The results showed that the molecular weight of recombinant protein pGEX-4T-1-MadCam-1 was 50 ku. Then rabbits were injected with purified protein to induce immunoreactions and the potency of the antibody was as high as 1 ∶ 32000. Therefore, the polyclonal antibody of MadCam-1 was induced. The results of this study lay a foundation for further study of the function of dairy cows MadCam-1.关键词
MadCam-1/克隆/表达/纯化/多克隆抗体Key words
MadCarn-1/ cloning/expression/purification / polyclonal antibody分类
生物学引用本文复制引用
刘文博,陈媛媛,孙玲,李德朋,付云贺,张乃生,郭梦尧,杨正涛..奶牛黏附分子MadCam-1基因的克隆及原核表达[J].中国畜牧兽医,2011,38(8):131-134,4.基金项目
国家自然科学基金资助项目(30972225). （30972225）
