通用黏膜免疫佐剂的制备

摘要

目的 采用同源重组技术制备通用黏膜佐剂.方法 将枯草芽胞杆菌CotB基因与融合基因AB连接后插入整合质粒pDG1662得到重组质粒pDG1662-CotB-AB,电转化法将线性化重组质粒转化枯草芽胞杆菌1A771,抗生素抗性筛选,PCR、免疫荧光法鉴定免疫球蛋白结合蛋白基因重组及蛋白表达情况.结果 AB基因成功重组入枯草芽胞杆菌基因组中,免疫球蛋白结合蛋白在芽胞表面得到了有效表达,并具有结合免疫球蛋白的生物活性.结论 本研究成功制备了黏膜免疫佐剂.%Objective To introduce a recombinant functional domains immunoglobulin binding protein(FDIBP) gene AB into the Bacillus subtilis genome by means of homologous recombination, and then to display FDIBP on the spore as adjuvant of mucosa vaccine. Methods A recombinant AB gene fused with Bacillus subtilis CotB gene was introduced into integration vector pDG1662 to generate plasmid pDG1662-CotB-AB. Bacillus subtilis 1A771 was transformed with linearized pDG1662-CotB-AB by electroporation. Recombinant strains were screened by antibiotic resistance and were identified by PCR and immunofluorescence microscopy. Results The AB gene integrate into Bacillus subtilis genome successfully and FDIBP was displayed on the of spore's surface. Conclusion The success of FDIBP expression on the spore's surface with affinity to hlgG is a major achievement toward constructing a adjuvant of mucosa vaccine.