小鼠Gli-similar-1(Glis1)基因新转录本功能分析  

Function Analysis of a New Isoform of Mouse(Mus musculus) Gli-similar-1(Glis1) Transcript

作　　者：王小海[1] 杨力侠[1] 吴勇延[1] 杜娟[1] 唐波[1] 郭泽坤[1] 

机构地区：[1]西北农林科技大学生命科学学院,杨凌712100

出　　处：《农业生物技术学报》2012年第7期815-821,共7页Journal of Agricultural Biotechnology

基　　金：国家自然科学基金项目(No.30870266;No.31172279)

摘　　要：Gli-similar-1(Glis1)是一种在小鼠卵母细胞和受精卵一、二细胞期高表达蛋白,在小鼠胚胎发育中起着重要的作用。本实验以小鼠(Mus musculus)肾为材料,克隆小鼠 Glis1 基因,并构建与绿色荧光蛋白融合表达的真核表达载体 pEGFP-C1-Glis1。研究了 Glis1 蛋白的表达和亚细胞定位,并利用 qPCR 检测过表达 Glis1 对 Oct4、Sox2、c-Myc、N-Myc、Klf4、Nanog、Nrgn 和 Tspan18 等基因表达的影响。结果表明,从小鼠肾中成功克隆到 Glis1 的一个新转录本(GenBank 登录号: JQ043365),其第三个核定位信号缺失 4 个氨基酸,C 端结构域缺失 124 个氨基酸,包括富含脯氨酸结构域全部氨基酸序列。Glis1 定位于细胞核,过表达能够显著上调Tspan18。研究结果表明,Glis1 新转录本与现有转录本亚细胞定位结果一致,其在小鼠胚胎发育和小鼠成纤维细胞重编程过程中可能发挥不同的功能。Gli-similar-1 （Glisl） is high expressed in unfertilized oocytes and in embryos at the 1, 2-cell stage, and it plays an important role on mouse embryos devolpment. Glisl gene was cloned from mouse （Mus musculus） kidney and constructed eukaryofic expression vector pEGFP-C1-Glisl. The expression and subcellular localization of pEGFP-C1-Glisl was detected by Western blot and GFP signal. In addition, the expression of several transcription factors was examined by quantitative RT-PCR after overexpression of Glis 1. These factors included Oct4, Sox2, c-Myc, N-Myc, Klf4, Nanog, Nrgn and Tspan18. The results showed that a new isoform of Glisl transcript （Genbank accession no. JQ043365） was cloned. This transcript encoded a truncated protein that lacked four amino acids in the third NLS （nuclear localization signal）, and lacked 124 amino acids at the C-terminus, including all amino acids in proline-rich region. This new Glisl was localized in the nucleus and overexpression of Glisl dramatically upregulates the mRNA level of Tspan18. The result indicated that subcellular localization of the new isoform of mouse Glisl transcript was identical with other existing transcripts, it might play a different role on mouse embryos devolpment and in induced pluripotent stem cells （iPSCs）.

关 键 词：Gli-similar-1(Glis1) twosl8 诱导性多能干细胞(iPSCs) 亚细胞定位 

分 类 号：Q954.432[生物学—动物学]