摘要
在研究水稻基因OsWAK1功能的过程中,利用荧光分子标记的方法,将OsWAK1与绿色荧光蛋白GFP构建为融合蛋白,利用注射法使表达OsWAK1::GFP融合蛋白的农杆菌浸入烟草叶片。通过荧光显微观察,根据烟草叶片表皮细胞中是否产生绿色荧光来判断融合蛋白是否表达。研究中同时利用了3个不同的烟草品种,利用同样的方法和操作程序,但是最后根据烟草叶片中绿色荧光的检测结果发现,不同的烟草品种对OsWAK1::GFP融合蛋白的表达差异很大。OsWAK1::GFP融合蛋白能够在心叶烟叶片中表达,却未能检测到其在本生烟和三生烟两个品种的叶片中表达。
In order to study the subcellular localization of rice OsWAK1, the fusion protein of OsWAKI::GFP (green fluorescence protein) was constructed. A Agrobacterium tumefaciens strain EHA 105 containing OsWAKI:: GFP fusion was inoculated through the stomata using a 1-ml plastic syringe and by gentle pressure on the lower epidermal surface of tobacco leaves. In the experiment three tobacco species Nicotiana glutinosa, Nicotiana tabacum cv. Sam and Nicotiana benthamiana respectively were used. As a result, the green fluorescence was detected from Nicotiana glutinosa and couldn' t, however, be detected from Nicotiana tabacum ev. Sam and Nicotiana bentharniana, which indicate the difference of transient expression of the fusion protein, the fusion protein was transiently expressed in Nicotiana glutinosa, but not in the other two species.
出处
《中国农学通报》
CSCD
北大核心
2009年第8期58-61,共4页
Chinese Agricultural Science Bulletin
基金
国家基础研究计划"农作物重大病害成灾的机理与控制的基础研究"(No.2006CB101905)