日本脑炎病毒C蛋白对其复制子载体自主复制活性的影响  

Influence of Japanese enciphalitis virus capsid protein on the self-replicate ability of JEV replicon vectors

作　　者：黄莺[1] 刘珊[2] 杨鹏[1] 王超[1] 杜韫[1] 孙志伟[1] 俞炜源[1] 

机构地区：[1]军事医学科学院生物工程研究所蛋白质工程研究室,北京100071 [2]中国人民解放军第二炮兵总医院病理科,北京100088

出　　处：《生物工程学报》2010年第8期1088-1094,共7页Chinese Journal of Biotechnology

摘　　要：为了研制具有高效自主复制能力的日本脑炎病毒(JEV)复制子载体,验证其作为新型复制子疫苗载体的可能性。以保留全长核心蛋白C基因的JEV复制子载体pCTCJEV为基础,通过PCR的方法减短C蛋白的部分基因序列,分别保留C23和C68位氨基酸,以LacZ基因作为报告基因,构建了C基因长短不同的JEV复制子载体pCMW-2M-1LACZ、pCMW-2M-3LACZ。将复制子载体转染表达JEV结构蛋白的细胞系CME-4,通过LacZ的表达检测JEV复制子载体表达外源蛋白的能力,反映了JEV的系列复制子载体的自主复制能力。结果保留C基因全长,C68、C23的复制子载体表达外源蛋白的能力相当,以上结果说明仅仅保留C蛋白的69个核苷酸即可保留JEV复制子载体的自主复制能力,为进一步优化JEV复制子载体,将该载体开发研制成为高效表达外源蛋白的疫苗载体提供了依据。To optimize a self-replicate Japanese enciphalitis virus （JEV） replicon, and to make it as an efficient vector to express the heterologous protein, we constructed three JEV replicons by PCR-based shortening the length of capsid genes. The vectors remained full or part of C gene, based on the JEV replicon pCTCJEV. Lac Z was selected as the reporter gene to verify the self-replicate ability of these DNA-based replicons. While transfected into the cell lines CME-4, which continuously expressing the JEV structure proteins C-prM-E, the JEV replicons pCMW-2M-1LACZ, pCMW-2M-3LACZ, which remained the first 23aa and 68aa of C protein, can express the reporter protein as the same level as pCMW-2M-LACZ with the full-length C protein. These results illustrated that the JEV replicon vector with 69-nt of the C gene can retain the self-replicate ability, and provide valuable tools to construct a possible vector for a long-lasting JEV RNA virus expression system.

关 键 词：JEV C蛋白 复制子载体 

分 类 号：R373[医药卫生—病原生物学]