表达绿色荧光蛋白的鸭肠炎病毒gE基因转移载体构建  

Construction of Duck Enteritis Virus gE Gene Transfer Vector Expressing Enhanced Green Fluorescent Protein

作　　者：乌伊罕[1] 赵妍[1] 刘晓玫[1] 张智慧[1] 马波[1] 王君伟[1] 

机构地区：[1]东北农业大学动物医学院,黑龙江哈尔滨150030

出　　处：《中国动物检疫》2009年第6期27-29,共3页China Animal Health Inspection

基　　金：国家"十一五"科技支撑计划;高致病性禽流感疫苗的研制与产业化;项目编号:2006BAD06A05-1

摘　　要：本实验利用PCR方法扩增DEVC-KCE株gE基因片段(包含gI基因及其侧翼序列),扩增产物克隆入pGEM-T载体测序后亚克隆至pUC19 EcoRI、SalI位点间,获得pUC-gE。将增强型绿色荧光蛋白表达盒插入pUC-gE BamHI位点,构建转移载体pUC-gE-EGFP。该转移载体有7个单一酶切位点可供外源基因插入,上下游侧翼分别为1.57Kb和1Kb。将转移载体pUC-gE-EGFP转染鸭胚成纤维细胞(DEF),观察到绿色荧光,说明EGFP基因获得有效表达,为开发以鸭肠炎病毒为载体的多价、多联基因工程疫苗提供了物质基础。The gE gene （including gI and its flanking sequences） was amplified by polymerase chain reaction （PCR） with extracted total DNA from DEV C-KCE strain as template and cloned into vector pGEM- T. Then the gE gene was subcloned into pUC19 vector digested by EeoR I and SalI, and a recombinant vector pUC-gE was constructed. At last the EGFP expression cassette was inserted into BamHI sites of the pUC-gE vector to obtain the DEV transfer vector pUC-gE-EGFP.There were 7 restriction sites for insertion of the foreign gene. The upstream and downstream flanking sequences were up to 1.57 kb and 1 kb. The transfer vector pUC-gE-EGFP was transfected into DEF cell, and the green fluorescence could be seen. The results showed that EGFP gene could be expressed correctly. It will be useful for developing the recombinant DEV expressing foreign gene（s）.

关 键 词：鸭肠炎病毒 转移载体 绿色荧光蛋白 

分 类 号：S852.659.1[农业科学—基础兽医学]