牛整合素β7亚基片段的表达及多抗制备

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 by)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.