中国兽医科学2011，Vol.41Issue(5)：479-483,5.
口蹄疫病毒3C基因与增强型绿色荧光蛋白基因的真核共表达
Co-expression of 3C gene of foot-and-mouth disease virus and enhanced green fluorescent protein gene in Eukaryote
摘要
Abstract
To provide an experimental basis for verifying siRNAs effect on inhibition of foot-and-mouth disease virus(FMDV) replication,3C gene was amplified by reverse transcription-polymerase chain reaction (RT-PCR) from Jiangsu/China/2005 strain of Asia 1 serotype of FMDV and was cloned into pEGFP-N1 expression vector with enhanced green fluorescent protein(EGFP) gene and the constructed recombinant was confirmed by PCR, restriction analysis and sequencing. PK-15 cells were transfected with positive recombinant plasmid,the expression of EGFP and transcription level of 3C gene were determined. Both PCR and restriction analysis demonstrated that the target gene fragment was corresponding to that expected in size. The sequence of the target gene was consistent with that of corresponding gene fragment in Jiangsu/ China/2005 strain. The expression of EGFP was observed by both fluorescent microscopy and flow cytometry. The transcription of 3C gene could be detected by real-time quantitative PCR in the transfected PK-15 cells. The recombinant plasmid for co-expression of FMDV 3C gene with EGFP gene was successfully constructed and expressed in PK-15 cells.关键词
口蹄疫病毒/3C基因/增强型绿色荧光蛋白/真核共表达Key words
foot-and-mouth disease virus/ 3C gene/ enhanced green fluorescent protein(EGFP)/ eukaryotic co-expression分类
农业科技引用本文复制引用
魏衍全,田宏,吴锦艳,陈妍,尚佑军,窦永喜,刘湘涛,才学鹏..口蹄疫病毒3C基因与增强型绿色荧光蛋白基因的真核共表达[J].中国兽医科学,2011,41(5):479-483,5.基金项目
国家"十一五"科技支撑计划项目(2006BAD06A03) （2006BAD06A03）
转基因生物新品种培育重大专项(2009ZX08008-010B) （2009ZX08008-010B）
