Crop Protection Division, National Academy of Agricultural Science, Rural Development Administration, Wanju 565-851, Republic of Korea.
Department of Agricultural Biotechnology and Plant Genomics and Breeding Institute, Seoul National University, Seoul 151-921, Republic of Korea.
Sci Rep. 2016 Feb 29;6:22436. doi: 10.1038/srep22436.
Transient gene expression approaches are valuable tools for rapid introduction of genes of interest and characterization of their functions in plants. Although agroinfiltration is the most effectively and routinely used method for transient expression of multiple genes in various plant species, this approach has been largely unsuccessful in soybean. In this study, we engineered soybean mosaic virus (SMV) as a dual-gene delivery vector to simultaneously deliver and express two genes in soybean cells. We further show the application of the SMV-based dual vector for a bimolecular fluorescence complementation assay to visualize in vivo protein-protein interactions in soybean and for a co-immunoprecipitation assay to identify cellular proteins interacting with SMV helper component protease. This approach provides a rapid and cost-effective tool for transient introduction of multiple traits into soybean and for in vivo characterization of the soybean cellular protein interaction network.
瞬时基因表达方法是快速引入目的基因并研究其在植物中功能的有用工具。尽管农杆菌浸润是在多种植物物种中瞬时表达多个基因最有效和常规使用的方法，但该方法在大豆中基本不成功。在这项研究中，我们设计了大豆花叶病毒 (SMV) 作为双基因传递载体，以在大豆细胞中同时传递和表达两个基因。我们进一步展示了基于 SMV 的双载体在大豆中进行双分子荧光互补测定以可视化体内蛋白质-蛋白质相互作用以及用于共免疫沉淀测定以鉴定与 SMV 辅助蛋白酶相互作用的细胞蛋白的应用。该方法为大豆中瞬时引入多种性状以及体内鉴定大豆细胞蛋白相互作用网络提供了快速且具有成本效益的工具。
