摘要
旨在构建一个适用于农杆菌的遗传转化载体,为研究农杆菌T-复合物招募蛋白VBP3的编码基因Atu4856的表达调控机制奠定基础。通过聚合酶链式反应(PCR)获得该编码基因5'端上游可能含有转录活性的DNA片段。用绿色荧光蛋白基因(gfp)和495 bp Atu 4856启动子构成嵌合基因,以绿色荧光蛋白基因(gfp)为报告基因,体外构建启动子探针prset-agfp1,转化大肠杆菌,通过抗生素筛选和对绿色荧光蛋白(GFP)的跟踪观察,最终确定有启动子转录活性的基因片段。结果表明,该载体适用于农杆菌T-复合物招募蛋白VBP3的编码基因Atu4856的表达调控机理的研究。
The aim of this study was to construct a genetic transformation vector suitable for Agrobacterium tumefaciens,and lay the foundation for the study on the regulation of the expression of Atu4856,which encodes the T-complex recruitment protein VBP3 in Agrobacterium tumefaciens.By polymerase chain reaction(PCR),the DNA fragment upstream of the 5'end of the gene may contain transcriptional activity.The green fluorescent protein gene(gfp)and 495 bp Atu4856 promoter were composed as chimeric genes,and the promoter probe vector prset-agfp1 with green fluorescent protein gene(gfp)as reporter gene was constructed in vitro and transformed into E.coli.Through antibiotic screening and tracking of green fluorescent protein(GFP),successful identification of a gene fragment with promoter transcriptional activity.The results indicated that this expression vector was suitable for the study of the regulation mechanism of the expression of Atu4856,the encoding gene of Agrobacterium T-complex recruiting protein VBP3.
作者
刘啸宇
后梦情
程安琪
胡传俊
姚坤
吴萧
LIU Xiaoyu;HOU Mengqing;CHENG Anqi;HU Chuanjun;YAO Kun;WU Xiao(College of Biology and Food Engineering,Suzhou University,Suzhou,Anhui 234200,China)
出处
《农产品加工》
2022年第11期18-20,25,共4页
Farm Products Processing
基金
2020年安徽省大学生创新创业计划训练项目(S202010379060)。
关键词
农杆菌
启动子
绿色荧光蛋白
原核表达载体
Agrobacterium tumefaciens
promoter
green fluorescent protein
prokaryotic expression vector
