问号赖型钩端螺旋体内鞭毛蛋白与外膜蛋白基因融合DNA疫苗载体的构建  

Construction of DNA Vaccine Including a Chimeric Gene Encoding Flagellin and Outer Membrane Protein Antigen from Leptospira Interrogons Serovar Lai

作　　者：王敏[1] 戴保民[1] 游自立[1] 方之茂[1] 王雅静[2] 

机构地区：[1]四川大学华西基础医学与法医学院钩体病研究室,成都610041 [2]四川大学华西基础医学与法医学院寄生虫学教研室

出　　处：《华西医科大学学报》2002年第2期169-171,共3页Journal of West China University of Medical Sciences

基　　金：国家自然科学基金资助 (批准号 39970 6 6 7)

摘　　要：目的　为增强问号赖型钩端螺旋体 (钩体 0 17株 ) DNA疫苗内鞭毛蛋白基因 (fla B2 )与外膜抗原基因 (omp L1 )的免疫保护作用 ,构建一种能表达 fla B2 与 om p L1 蛋白融合蛋白的 DNA疫苗载体。方法　通过聚合酶链反应分别扩增 0 17钩体 fla B2 与 omp L1 片段并进行融合 ,获得的嵌合基因 om p L1 - fla B2 中含有 10个氨其酸的中间接头序列 ,以维持其空间构象。结果　经酶切鉴定 ,证实有一 1.8kb片段插入载体 ,其 fla B2 及 om p L1 DNA测序结果与文献报道的一致。结论　表达 0 17株钩体 fla B2 与 om p L1 抗原融合蛋白的Objective To construct a fusion expression vector for DNA vaccine, including a flagellin gene (flaB 2) and an outer membrane protein gene (ompL 1) to enhance the protective immunity of Leptospira interrogans serova Lai (Strain 017). Methods The DNA fragments encoding flaB 2 and ompL 1 were amplified respectively by PCR and then fused. The obtained chimeric gene ompL 1-flaB 2 contained a synthetic linker of ten amino acids to sustain its space conformation. Results Identified by restriction enzyme analysis,an insert fragment of 1.8kb was demonstrated. FlaB 2 and ompL 1 respectively had the same sequence as reported by DNA sequencing analysis. Conclusion A fusion expression plasmid containing flaB 2 and ompL 1 was constructed.

关 键 词：外膜蛋白 基因融合 DNA疫苗 载体 构建 问号赖型钩端螺旋体 内鞭毛蛋白 

分 类 号：R392[医药卫生—免疫学]