野大麦CIPK基因的亚细胞定位

The subcellular localization of CIPK from Hordeum brevisubulatum

为明确野大麦CIPK基因在植物亚细胞水平上的定位情况,以已知细胞质膜定位的拟南芥AtCBL1基因和液泡膜定位的AtCBL3基因作为参照,构建了这两个基因分别与红色荧光蛋白基因(RFP)融合的植物表达载体(参照载体),以及野大麦CIPK基因与绿色荧光蛋白基因(GFP)融合的植物表达载体(目标载体),利用基因枪转化法将参照载体和目标载体共转化洋葱上表皮细胞,瞬时表达后利用激光共聚焦显微镜进行共定位分析。结果表明,该基因主要定位于细胞膜和细胞核,为进一步深入分析该基因的功能提供了重要依据。

In order to determine the subcellular localization of CIPK from Hordeum brevisubulatum,using Arabidopsis plasma membrane-localized AtCBL1 and vacuole membrane-localized AtCBL3 as marker genes, two plant expression vectors were constructed as reference vectors, in which these two marker genes were fused with RFP, respectively. The target vector was constructed harbouring CIPK from Hordeum brevisubulatum fused with GFP. The reference and target vectors were co-transformed into onion epidermal ceils by particle bombardment method. The fluorescent sigals were observed by Confocal laser scanning microscope. Transient expression showed that CIPK from Hordeum brevisubulatum was localized at the plasma membrane as well as nucleus. The results provided an important basis for futher analyses of the function of CIPK from Hordeum brevisubulatum.