制备肠道病毒71型黏膜疫苗的初步探索

• 作 者: 李志会 岳盈盈 宋楠楠 李鹏 孟红
• 机 构: 山东省医学科学院基础医学研究所山东省罕少见病重点实验室,
• 出 处: 山东大学学报(医学版)
• 发表日期: 2012年02期
• 关键词: 肠道病毒71型 病毒结构蛋白VP1 黏膜疫苗 枯草芽胞杆菌芽胞

目的利用同源重组技术将肠道病毒71型衣壳蛋白VP1基因重组入枯草芽胞杆菌基因组,进而将VP1蛋白展示在其芽胞表面制备EV71黏膜疫苗。方法将枯草芽胞杆菌CotB基因与VP1基因连接后插入整合质粒pDG1662,得到重组质粒pDG1662-CotB-VP1,电转化法转化枯草芽胞杆菌1A771,抗生素抗性筛选,PCR、Westernblot、免疫荧光鉴定VP1基因重组及蛋白表达情况。结果 VP1基因成功重组入枯草芽胞杆菌基因组中,VP1蛋白在芽胞表面得到了有效表达。结论 EV71衣壳蛋白在枯草芽胞杆菌芽胞表面得到了有效表达,为研究EV71黏膜疫苗奠定基础。

OBJECTIVE: To construct the EV71 mucosal vaccine by recombining VP1 gene of enterovirus 71 capsid protein into Bacillus subtilis genome by homologous recombination technology. Methods The recombinant plasmid pDG1662-CotB-VP1 was inserted into the plasmid pDG1662 after the CotB gene of Bacillus subtilis was ligated with the VP1 gene. The recombinant plasmid was transformed into Bacillus subtilis 1A771 by electroporation. The antibiotic resistance screening, PCR and Westernblot were used to identify the recombinant VP1 gene And protein expression. Results The VP1 gene was successfully recombined into Bacillus subtilis genome and the VP1 protein was efficiently expressed on the spore surface. Conclusion EV71 capsid protein is efficiently expressed on the surface of Bacillus subtilis spores, which lays the foundation for the study of EV71 mucosal vaccine.