牛整合素β7亚基片段的表达及多抗制备  

Expression and Polyclonal Antibodies Preparation of the Fragment of β7 Subunit of Bovine Integrin

作　　者：王海霞[1] 杨正涛[1] 陈媛媛[1] 刘文博[1] 张雯[1] 张乃生[1] 

机构地区：[1]吉林大学畜牧兽医学院,吉林长春130062

出　　处：《中国畜牧兽医》2011年第3期100-103,共4页China Animal Husbandry & Veterinary Medicine

基　　金：国家自然科学基金资助项目(30972225)

摘　　要：为制备牛整合素β7亚基片段的多克隆抗体,根据GenBank中牛整合素β7基因序列,合成了β7亚基288 bp(140-427 bp)的cDNA片段,通过DNA重组法构建表达质粒,在大肠杆菌中经IPTG诱导表达。表达产物经Western blotting鉴定,结果证实成功表达了分子质量约为15 ku的β7蛋白。超声破碎细菌后,Ni-NTA树脂进行纯化,免疫兔子获得多抗。间接ELISA检测多抗效价为1∶32000,为进一步研究牛整合素α4β7的功能奠定基础。In order to prepare polyclonal antibodies of the fragment of bovine integrin β7 subunit.According to the bovine integrin β7 nucleotide sequence reported in the GenBank,the fragment of β7 subunit was synthetized.Expression vector was constructed by using DNA recombination technology.After being induced by IPTG,the fragment of β7 was expressed in E.coli.The expression product was identified by Western blotting.Results showed that the fragment of β7 with relative molecular mass of 15 ku was expressed successfully in E.coli.The expression product was purified by Ni-NTA resin.Then,the polyclonal antibody was prepared by inoculating rabbits with β7 purified product.The ELISA titer of the polyclonal antibody was approximately 1∶32000.The results of this study lay a foundation for further study of the function of bovine integrin α4β7.

关 键 词：整合素β7 原核表达 多抗 

分 类 号：Q51[生物学—生物化学]