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[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,对其亚细胞定为进行研究。[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] To determine the localization of OsWRKY78 protein in plants. [Method] According to the complete sequence of OsWRKY78 in GenBank, primers were designed and OsWRKY78 was amplified by RT-PCR. OsWRKY78 gene was cloned and recombined with plasmid pBinGFP containing green fluorescent protein (GFP) gene. Bacterial PCR and restriction enzyme digestion were performed. Finally, the recombinant vector was transformed into Arabidopsis by Agrobacterium tumefaciens-mediated bud soaking, and its subcellular localization was studied. [Result] The recombinant plasmid pBinGFP-OsWRKY78 was obtained by the test cloning. The colony PCR and restriction enzyme digestion showed that the constructed expression vector was correct and transformed into Arabidopsis. The transgenic plants were obtained. Fluorescence microscopy results showed that the expression product of OsWRKY78 Mainly located in the nucleus. [Conclusion] The results of this study lay a foundation for further study on the function of OsWRKY78 gene and its role in related signal transduction, and provided the theoretical basis for further study on the relationship between OsWRKY78 gene and N. lugens.