【作者】 郭恒； 刘娟； 李慧萍； 王学林； 孙树民； 张茂林； 高丽芳； 徐德启； 刘明远；

【Author】 GUO Heng~1,LIU Juan~1,LI Hui-ping~1,WANG Xue-lin~1, SUN Shu-min~1,ZHANG Mao-lin~1,GAO Li-fang~1,XU De-qi~(1,2),LIU Ming-yuan~1 (1.Key Laboratory for Zoonosis Research,Ministry of Education,Institute of Zoonosis,Jilin University,Changchun, Jilin,130062,China,2.Division of Bacterial,Parasitic and Allergenic Products.FDA-CBER,USA MD 20892)

【机构】 人兽共患病研究教育部重点实验室,吉林大学人兽共患病研究所； 美国食品与药品管理局(FDA)；

【摘要】 为验证冰核蛋白表面展示系统的可行性与优越性,将编码狂犬病毒(Rabies virus,RV)主要免疫保护性抗原糖蛋白G基因片段融合到丁香假单胞菌冰核蛋白截断的N端,构建表面展示载体pET28a-INP-RVG。转化大肠埃希菌BL21(DE3)后16℃低温诱导表达,SDS-PAGE检测表明在约78 ku和56 ku处有明细的蛋白带出现,大小与理论值相符。完整细胞ELISA实验结果表明,重组蛋白在大肠埃希菌表面成功展示。为进一步研制基于沙门菌的多抗原表位展示性重组疫苗、新型狂犬病疫苗奠定基础。更多还原

【Abstract】 The gene encoding Rabies virus(RV)glycoprotein(G protein) was fused to the C terminus of ice nucleation protein(INP),an outer membrane protein of Pseudomonas syringae.The fragment was cloned into prokaryotic expression vector pET28a(+ ),and a recombinant vector pET28a-INP-RVG was constructed. After transforming into Escherichia coli BL21(DE3),the recombinant strain was induced by 1 mM IPTG at 16℃and about 78 ku protein was detected with SDS-PAGE.The result of Western-blot indicated that the expressed protein possessed the antigenic specificity which could be recognized by rabbit anti-RV serum.The indirect immunofluorescent test and whole-cell enzyme linked immunosorbent assay showed that the expressed protein was secreted on the cell surface of Escherichia coli.The recombinant vector is background for the studies on recombinant vaccines of displaying several epitopes and a new type vaccine against rabies by using attenuated salmonella as vector.更多还原