表达流感病毒神经氨酸酶基因的重组腺病毒的构建  

Construction of Recombinant Adenovirus Expressing Influenza virus Neuraminidase Gene

作　　者：赵小东[1] 韩峰[1] 王艳[1] 杨贵宾[1] 应革[1] 林勇平[2] 王璇[2] 吴淑华[1] 

机构地区：[1]中国疾病预防控制中心病毒病预防控制所病毒基因工程国家重点实验室,北京100052 [2]广州医学院,广州510182

出　　处：《中国病毒学》2003年第3期213-216,共4页Virologica Sinica

摘　　要：通过RT-PCR方法扩增流感病毒神经氨酸酶基因,将其克隆到腺病毒穿梭载体pTrackCMV,此重组质粒与腺病毒DNA共转化大肠杆菌BJ5183,通过细菌内同源重组获得重组腺病毒DNA,将其转染293细胞获得重组腺病毒。经PCR证实目的基因已整合至腺病毒基因组中,western blot检测到神经氨酸酶的表达。重组病毒经滴鼻和灌胃两种途径免疫小鼠,结果表明2次免疫后滴鼻组和灌胃组均产生明显的免疫应答反应,滴鼻组的免疫效果优于灌胃组。The neuraminidase (NA) gene of Influenza virus A/FM/1/47 was amplified by RT-PCR and cloned into adenovirus vector pTrack-CMV. The recombinant plasmid and adenovirus DNA was cotransformated into E.coli BJ5183 and the recombinant adenovirus genomic DNA was obtained through homological recombination. The recombinant adenovirus was gained after transfecting 293 cell line with the genomic DNA. The intergration of NA in resultant adenovirus and the expression of NA gene were confirmed by PCR and western-blot respectively. BALB/c mice were immunized intranasally and orally respectively. ELISA result showed that systemic immue responses to Influenza virus could be induced effectively in both intranasal and oral route. The immunization efficacy of intranasal route was superior to that of oral route.

关 键 词：流感病毒 神经氨酸酶基因 重组腺病毒 疫苗 

分 类 号：R373[医药卫生—病原生物学]