中国兽医科学2011，Vol.41Issue(1)：48-52,5.
山羊痘病毒RPO30基因绿色荧光蛋白质粒的构建及融合表达
Construction of recombinant plasmid for fusion expression of goatpox virus RPO30 gene and enhanced green fluorescent protein gene
摘要
Abstract
The aim of this study was to construct a recombinant plasmid for fusion expression of goatpox virus(GPV) RPO30 gene and enhanced green fluorescent protein(EGFP) gene and express it in BHK-21 cells.The RPO30 gene was amplified from GPV Gulang strain by PCR and inserted into the expression vector pEGFP-N1 containing EGFP gene, and the constructed recombinant plasmid was identified by restriction enzyme digestion analysis and sequencing.BHK-21 cells were transfected with screened positive recombinants and the expression of EGFP and transcription level of RPO30 gene were determined.Both restriction enzyme digestion analysis and sequencing proved that the target gene fragment was consistent with that expected in size.The sequence of the target gene was identical to that of corresponding gene fragment in Gulang strain.The expression of EGFP was observed by both fluorescent microscopy and flow cytometry.RT-PCR proved the transcription of RPO30 gene in the transfected BHK-21 cells.The recombinant plasmid for fusion expression of GPV RPO30 gene and EGFP gene was successfully constructed and the genes were expressed in BHK-21 cells.关键词
山羊痘病毒/RPO30基因/增强型绿色荧光蛋白/融合表达分类
农业科技引用本文复制引用
赵志荀,吴国华,颜新敏,崔力凡,李健,朱海霞,张强..山羊痘病毒RPO30基因绿色荧光蛋白质粒的构建及融合表达[J].中国兽医科学,2011,41(1):48-52,5.基金项目
农业部转基因生物新品种培育重大专项(2009ZX08008-010B) （2009ZX08008-010B）
甘肃省重大科技专项(092NKU032) （092NKU032）
