柯萨奇B组病毒VP1胞质表达系统的建立  

Cytoplasmic expression of VP1 gene of Coxsackievirus B3

作　　者：陈洪[1] 刘晶星[1] 陈淑云[1] 何平[1] 胡宝瑜[1] 李振红[1] 

机构地区：[1]上海第二医科大学病原生物学教研室,200025

出　　处：《中华实验和临床病毒学杂志》2005年第1期46-48,共3页Chinese Journal of Experimental and Clinical Virology

基　　金：国家自然科学基金资助项目 (3 9970 691)

摘　　要：目的　利用T7RNA聚合酶介导的胞质表达系统增加目的抗原的表达量 ,提高基因疫苗的免疫效果。方法　 (1)构建带有T7启动子、脑心肌炎病毒 (EMCV) 5′非编码区和柯萨奇B组病毒衣壳蛋白VP1的质粒pT7EMCVP1。将该质粒和编码T7RNA酶的pAR 3132共转染HeLa细胞和小鼠腹腔巨噬细胞 ;(2 )将上述质粒分别转化减毒鼠伤寒沙门菌SL72 0 7,让带有不同质粒的两种载体细菌共感染小鼠腹腔巨噬细胞。结果　 (1)经共转染 ,在HeLa细胞和小鼠腹腔巨噬细胞中 ,目的抗原VP1在胞质中的表达比单独转染真核表达质粒pcDNA3VP1增加 2～ 4倍 ;(2 )两种质粒载体细菌感染小鼠巨噬细胞后 ,目的抗原VP1也能在细胞中较好表达。结论　pT7EMCVP1和pAR 3132能在HeLa细胞和小鼠腹腔巨噬细胞胞质中表达 。Objective To increase the immune effect of gene vaccine, T7 RNA polymerase was used to establish a system of cytoplasmic expression. Methods (1) The plasmid pT7 EMCVP1, including T7 promoter sequence, 5′-untranslated sequence of encephalomyocarditis (EMC) virus, VP1 sequence of coxsackievirus B3 (CVB3), was cotransfected with the plasmid pAR 3132, which codes for the T7 RNA polymerase, into Hela cells and murine peritoneal macrophages. (2) The plasmid pT7 EMCVP1 and pAR 3132 were respectively transformed into the attenuated Salmonella typhimurium SL 7207. The two kinds of transformed bacteria were coinfected into murine peritoneal macrophages. Results (1) The target antigen VP1 in the cytoplasm was about 2-4-fold higher than that of pcDNA3 VP1 singly transfected. (2) After the murine peritoneal macrophages were coinfected by two kinds of transformed bacteria, the target antigen VP1 could also be detected. Conclusion The pT7 EMCVP1 and pAR 3132 could be expressed in the cytoplasm of HeLa cells and murine peritoneal macrophages and the amount of the antigen VP1 increased remartally as compared with that of pcDNA3 VP1 singly transfected.

关 键 词：VP1 小鼠腹腔巨噬细胞 柯萨奇B组病毒 HELA细胞 抗原 质粒 共转染 胞质 种质 表达量 

分 类 号：R346[医药卫生—基础医学]