牛传染性鼻气管炎病毒gD基因的克隆与表达

Cloning and Expression of Infectious Bovine Rihinotracheitis Virus gD Gene

构建pET32a重组表达载体,转化到BL21(DE3),诱导表达牛传染性鼻气管炎病毒重组gD蛋白。用已发表的IBRV的gD基因序列设计一对特异性引物,通过PCR方法扩增一条766bp的目的基因gD,将gD基因克隆到原核表达载体pet32a,得到重组表达载体pet32-gD,转化到BL21(DE)中,通过IPTG诱导获得融合蛋白。重组表达蛋白经纯化后,免疫印迹分析。结论证明表达的重组蛋白具有良好的免疫原性。

pET32a recombinant expression vector was constructed and transformed into E.coli BL21 to induce the expression of recombinant infectious bovine rhinotracheitis virus(IBRV) gD protein.According to the sequence of IBRV gD gene reported ,a pair of PCR primers were designed .Then PCR was carried out to amplify an objective fragment of 766 bp.The PCR products were cloned into expression vector pET32a and the recombinant plasmids pet32-gD were abtained.Then, pET 32-gD was transformed into the competent E.coli BL21. Recombinant gD protein induced by IPTG was expressed ,purified and analyzed by western bloting. The results demonstrated that the recombinant protein could react with the antibody against IBRV by western blot,with the good antigenicity.