论文部分内容阅读
在克隆水稻编码磷脂酰丝氨酸合酶的短穗颈基因SUI1的基础上,利用Gateway方法分别构建了N端和C端两种GFP融合载体pMDC45-SOVT和pMDC201-SOV。利用农杆菌侵染烟草叶片的方法,观察SUI1蛋白在烟草叶肉细胞中的瞬时表达情况,结果显示GFP-SUI1定位于质膜和核上,而SUI1-GFP定位于质膜和核膜上,并且两种融合蛋白在质膜上的分布都是不连续的。由于N端融合可能改变了目标蛋白的折叠方式及跨膜方式,从而影响蛋白的正确定位,因而SUI1蛋白主要定位于烟草叶肉细胞的质膜和核膜中。
Based on the cloned short panicle gene SUI1 encoding phosphatidylserine synthase in rice, two GFP fusion vectors pMDC45-SOVT and pMDC201-SOV were constructed by Gateway method. The transient expression of SUI1 protein in tobacco mesophyll cells was observed by Agrobacterium tumefaciens infection. The results showed that GFP-SUI1 localized on the plasma membrane and nucleus, while SUI1-GFP localized on the plasma membrane and nuclear membrane, and The distribution of the two fusion proteins on the plasma membrane is discontinuous. Since the N-terminal fusion may change the folding mode and the transmembrane mode of the target protein, thereby affecting the correct localization of the protein, the SUI1 protein mainly locates in the plasma membrane and the nuclear membrane of tobacco mesophyll cells.