论文部分内容阅读
目的 HPV16L1重组蛋白的表达为研制HPV16L1预防性蛋白疫苗及诊断试剂盒打下基础。方法 pPIC3 5 HPV16L1 GS115阳性菌株 ,经 0 5 %甲醇诱导 ,运用SDS PAGE电泳检测L1蛋白 ;用鼠抗HPV16L1单克隆抗体 ,经WesternBlotting检测蛋白的特异性 ;在透射电镜下观察类病毒颗粒 (VLPs)。结果 SDS PAGE检测结果表明 ,在5 5KD处有诱导蛋白带的出现 ;经WesternBlotting验证 ,该蛋白能与HPV16L1单克隆抗体特异结合 ,证明该蛋白确为HPV16L1蛋白 ;电镜下见到了直径约为 5 0nm、形态结构与天然病毒颗粒极其相似的VLPs。结论 pAIC3 5 HPV16L1重组质粒在甲醇营养型酵母菌中可经甲醇诱导产生HPV16L1晚期蛋白 ,该蛋白在酵母菌中可自我组装成VLPs。
Objective To lay a foundation for the development of HPV16L1 prophylactic protein vaccine and diagnostic kit. Methods The pPIC3 5 HPV16L1 GS115 positive strains were induced by 0.5% methanol, and the L1 protein was detected by SDS PAGE electrophoresis. The specificity of the protein was detected by Western blotting with the mouse anti-HPV16L1 monoclonal antibody. The virus-like particles (VLPs) were observed under transmission electron microscope. . Results The results of SDS PAGE showed that the inducible protein band appeared at 55KD. The Western blotting showed that the protein could specifically bind to the HPV16L1 monoclonal antibody, which proved that the protein was HPV16L1 protein. The diameter of the protein was about 50 nm under electron microscope. VLPs with morphological structures that are very similar to native virus particles. Conclusion The recombinant plasmid pAIC3 5 HPV16L1 can induce HPV16L1 late protein in methanol-producing yeast, which can self-assemble into VLPs in yeast.