摘要
以潮霉素抗性基因hph为筛选标记,以双元载体pPK2为基本骨架,将增强型绿色荧光蛋白基因egfp置于组成型启动子PgpdA和色氨酸C终止子TtrpC之间,构建了组成性表达egfp的载体pPK2-EGFP,利用根癌农杆菌介导的遗传转化法,转入贵阳腐霉(Pythium guiyangense)表达。对转化子进行RT-PCR和荧光显微镜检测,结果表明,egfp基因在贵阳腐霉转化子中能稳定表达。
A expression vector pPK2-EGFP containing a modified green fluorescent protein(egfp) gene was constructed using binary vector pPK2 as basic framework,in which the egfp gene was placed between promoter PgpdA and terminator TtrpC.The recombinant strains were obtained by Agrobacterium-mediated transformation system with Pythium guiyangense mycelia as acceptor using pPK2-EGFP vector containing hygromycin B resistance gene as marker.The results of RT-PCR analysis and flurescent microscopy showed that the egfp gene was stably expressed in P.guiyangense transformant.
出处
《湖北农业科学》
北大核心
2012年第24期5801-5803,5816,共4页
Hubei Agricultural Sciences
基金
贵州省科学技术基金项目(黔科合J字[2008]2084号)
