不同启动子表达载体的构建及其体外活性分析  

The Construction of Different Promoter Expression Vector and its Activity Analysis in vitro

作　　者：刘正伟[1] 李慧敏[1] 黄妙容[2] 郭凯[1] 王建丽[1] 李延鹏[2] 陈瑞爱[1,2] 

机构地区：[1]华南农业大学,广东广州510642 [2]广东大华农动物保健品股份有限公司,广东云浮527400

出　　处：《中国畜牧兽医》2014年第2期11-15,共5页China Animal Husbandry & Veterinary Medicine

摘　　要：本研究以禽网状内皮增生症病毒(reticuloendotheliosis virus,REV)囊膜糖蛋白(envelope,env)基因作为报告基因,定量检测不同启动子在体外的转录活性,为研发马立克氏病病毒(Marek’s disease virus,MDV)活载体疫苗、选择适宜的启动子提供依据。本试验构建6个含不同启动子,且能表达env基因的重组质粒,并将构建好的重组质粒转染到鸡胚成纤维细胞中,通过间接免疫荧光试验和实时荧光定量PCR检测不同启动子的表达转录水平。研究证明6种启动子均能使env基因在体外获得表达,且不同启动子的转录活性不同。比较发现MDV自身启动子相对其他组启动子的转录活性较弱,pec启动子的转录活性最强。The envelope （env） gene of reticuloendotheliosis virus （REV） as a report gene,could be used for quantitative de- tection of different promoters transcription activity in vitro, and provide the basis for choosing suitable promoter of living vec- tor vaccine of Marek＇s disease virus （MDV）. We constructed six recombinant plasmids,containing different promoter and ex- pressing env gene, transfected into chicken embryo fibroblast, detected different promoter transcriptional and expressional level by indirect immunofluorescence test and fluorescent quantitative PCR. The results showed that different promoters had differ- ent activity and all expressed env gene in vitro. Comparing to the other promoters, we found the own promoter activity of MDV was weak, the pec was the strongest in vitro.

关 键 词：禽网状内皮增生症病毒 囊膜糖蛋白基因 启动子 活性分析 

分 类 号：Q78[生物学—分子生物学]