牛乳头瘤病毒2型L1基因的克隆及原核表达

Cloning and expression of L1 gene of bovine papillomavirus type 2

为表达牛乳头瘤病毒2型(BPV2)衣壳蛋白L1基因,本研究采用PCR方法从奶牛皮肤肿瘤病料样品中扩增BPV2沙湾分离株L1基因,克隆至pMD18-T载体中,测序后进行分析。将BPV2沙湾株L1基因亚克隆于原核表达质粒pGEX4T-1中,经IPTG诱导L1蛋白表达,并对重组蛋白进行SDS-PAGE和western blot分析。SDS-PAGE分析显示,诱导后能够表达分子量80 ku的重组蛋白;western blot显示该蛋白可以与抗BPV2多克隆抗体发生反应。小鼠免疫试验表明,BPV2 L1重组蛋白可以诱导小鼠产生抗BPV2 L1特异性抗体;琼脂扩散试验显示,重组L1蛋白具有良好的免疫原性。本研究原核表达BPV2 L1蛋白,并证明其具有良好的反应原性及免疫原性,为牛乳头瘤病的疫苗和诊断试剂研发奠定了基础。

For expressing L1 gene of bovine papillomavirus type 2 （BPV2）, the BPV2 L1 gene was amplified from the skin lesions of affected cow by PCR and cloned into pGEX4T-1 vector for expressing in E.coli. The SDS-PAGE analysis revealed that the expressed recombinant L1 protein was 80 ku, which was recognized by anti-BPV2 polyclonal antibody detected by western blot. Immunologic tests showed that the recombinant L1 protein was able to induce specific antibodies agaist BPV2 in mice. Agar diffusion test demonstrated that the recombinant L1 protein could be used as antigen of diagnostic reagent for detections of BPV2 in infected cattle.