TRIM基因家族一个新成员的克隆及功能研究

The cloning and functional analysis of a novel TRIM protein

目的:克隆小鼠早期胚胎发育相关的新基因uTRIM,研究其胞内亚定位及功能。方法:通过数字差异显示搜索到可能和小鼠早期胚胎发育相关的新基因uTRIM,用RT-PCR方法从体外培养的小鼠囊胚中分离得到其全长cDNA;将其克隆到绿色荧光蛋白表达载体pEGFP-N1中,在HEK293细胞内表达后确定uTRIM分子的亚细胞定位。用RNAi对其在胚胎发育早期的功能进行初步的研究。结果与结论:uTRIM基因只在胚胎发育早期和成体睾丸组织中表达,亚细胞定位显示该蛋白位于细胞质中。RNAi结果显示抑制uTRIM表达使小鼠受精卵体外发育速度加快,提示uTRIM参与早期胚胎发育的负调控。

Objective： To clone a novel member of the mouse TRIM gene family, and investigate its subcellular location and function during the period of early embryonic development. Methods： A novel TRIM gene （uTRIM） was selected with digital difference display method, and its cDNA was amplified by RT-PCR with mouse embryonic mRNA as template. To identify the subcellular location of the uTRIM, its cDNA was subcloned into pEGFP-N1 vector and expressed in HEK 293 cells. RNAi was used to investigate the effect of uTRIM protein on the development of mouse embryo. Results and Conclusion. The cDNA of mouse uTRIM was cloned, RT-PCR results showed that the uTRIM gene was only expressed in adult testis and early stage embryo. RNAi results indicated that uTRIM may negatively regulate the development of mouse early embryo.