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建立携带荧光未分化标记物的小鼠胚胎干细胞系。

Establishing mouse embryonic stem cell line carrying a fluorescent undifferentiated marker.

作者信息

Teng Lu, Cheng Jun-Ying, Yang Yang, Zhang Chong-Ben

机构信息

College of life sciences, Peking University, Beijing 100871, China.

出版信息

Yi Chuan Xue Bao. 2004 Oct;31(10):1061-5.

Abstract

To label mouse ES cells,a cell line derived from the inner cell mass of 3.5-day blastocysts,with enhanced green fluorescent protein (EGFP), the vector of pRex-1-EGFP was transferred into mouse ES cells by electroporation. The expressions of Rex-1 in undifferentiated and differentiated ES cells were detected by the microscopic observation of EGFP and by RT-PCR. The results showed that the EGFP gene was transferred into the mouse ES cell line, and the transfected cells in undifferentiated state showed high levels of EGFP expression. When the cells began to differentiate, the EGFP expressions were gradually reduced. A mouse ES cell line expressing EGFP under the control of Rex-1 gene promoter was generated. The cell line provides a powerful approach for the research of the process of mammalian development and for the screening of small molecules that can regulate this process.

摘要

为了用增强型绿色荧光蛋白(EGFP)标记从小鼠3.5天囊胚内细胞团衍生而来的小鼠胚胎干细胞系,通过电穿孔将pRex-1-EGFP载体导入小鼠胚胎干细胞。通过EGFP的显微镜观察和RT-PCR检测未分化和分化的胚胎干细胞中Rex-1的表达。结果表明,EGFP基因已转入小鼠胚胎干细胞系,未分化状态的转染细胞显示出高水平的EGFP表达。当细胞开始分化时,EGFP表达逐渐降低。构建了一个在Rex-1基因启动子控制下表达EGFP的小鼠胚胎干细胞系。该细胞系为研究哺乳动物发育过程以及筛选可调节该过程的小分子提供了有力手段。

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