作　　者： ; ; ; ;

机构地区： 广州大学生命科学学院

出　　处： 《安徽农业科学》 2012年第19期9982-9984,10026,共4页

摘　　要： [目的]确定OsWRKY78蛋白在植物中的定位。[方法]根据GenBank数据库中OsWRKY78全序列设计引物,进行OsWRKY78的RT-PCR扩增,克隆了OsWRKY78基因,将该片段与带绿色荧光蛋白(GFP)基因的质粒载体pBinGFP重组,并对重组载体进行菌液PCR和酶切验证,最后利用农杆菌介导的花蕾浸泡法将重组载体转化到拟南芥中,对其亚细胞定位进行研究。[结果]试验克隆得到了pBinGFP-OsWRKY78重组载体,经菌落PCR与酶切检测表明构建的表达载体正确,其转化到拟南芥中后得到了转基因植株,荧光显微镜检测结果表明,OsWRKY78基因表达产物主要定位在细胞核中。[结论]该研究结果为深入研究OsWRKY78基因的功能及其在相关信号传导中的作用奠定了基础,也为进一步研究OsWRKY78基因与褐飞虱之间的关系提供了理论依据。 [Objective] The objective of the study was to understand the subcellular localization of OsWRKY78 protein in plants.[Method] In doing that,primers of OsWRKY78 gene were designed according to the OsWRKY78 full length sequence in Genbank.The gene was cloned by RT-PCR method.The gene was then recombined into a plasmid expression vector carrying green fluorescent protein（GFP） gene,pBinGFP.The recombinant was confirmed by PCR enzyme digestion.The recombinant plasmid pBinGFP-OsWRKY was transformed into Arabidopsis through Agrobacterium tumefaciens strain GV3101 and transgenic plant was obtained.[Result] Measured by fluorescence microscopy,the expression of OsWRKY78 and GFP fusion protein in root tip cells was localized in the nucleus.[Conclusion] This study laid the foundation for further investigating the function of OsWRKY78 gene and its role in related signal transduction and provided theoretical basis for exploring the relation between OsWRKY78 gene and brown planthoppers.

关 键 词： 基因 绿色荧光蛋白基因 表达载体 亚细胞定位

领　　域： [农业科学]