转GFP::cDNA基因拟南芥筛选及相关基因的功能分析

本研究以随机GFP::cDNA融合基因转基因拟南芥为材料,筛选到在细胞核或在细胞核和细胞质中均表达GFP信号的转基因株系58个。对这些转基因株系中的cDNA插入片段进行克隆,获得4株插入片段能按原初编码框进行编码的转基因株系。对插入片段为编码富含甘氨酸蛋白AtGRP8 C-末端(富含甘氨酸的结构域)的转基因株系R2的表型分析发现,连续白光、红光或蓝光下其幼苗的下胚轴比野生型的要短,且较低光照强度白光(低于100μmol m-2s-1)、蓝光(低于75μmol m-2s-1)下的差异更加明显,但是黑暗中其幼苗的下胚轴与野生型相比无明显差异,表明AtGRP8蛋白可能通过其C-末端功能域参与调控拟南芥的光形态建成反应。

Screening Transgenic Arabidopsis Expressing GFP:: cDNA Fusions and Functional Analysis of Related Gene

Transgenic Arabidopsis expressing random GFP：：cDNA fusions was used in this study. 58 Basta-resistant transgenic lines which expressing GFP： ：cDNA fusions in nuclei or in both nuclei and cytoplasm were obtained. The cD- NA fragment inserted into the transgenic plants were cloned and analyzed, and only four lines which express an in-frame fusion were obtained. Phynotype of one transgenic line R2, which expressing an in-frame fusion to a Gly-rich carboxy- terminal domain of AtGRP8 protein, was analyzed. The hypocotyl of R2 seedlings were shorter than that of wild type col- 4 grown in continuous white, blue or red light, especially under relative lower fluence rate white （less than 100 μmol m-2 s-1 ） or blue （ less than 75 μmol m-2 s-1 ） light, but there was no difference in the hypocotyl length when grown in the dark. The results indicated that AtGRP8 may be involved in controlling the photomorphogenesis of Arabidopsis seedling through its Gly-rich carboxy-terminal domain.