Zhou Jiang, Cheng Xuan, Lu Ya-Xin, Huang Cui-Fen, Yang Xiao
Genetic Laboratory of Development and Diseases, Institute of Biotechnology, Beijing 100071, China.
Sheng Wu Gong Cheng Xue Bao. 2002 May;18(3):286-90.
The transgenic mice that express Cre recombinase in a tissue specific manner is a powerful tool in generating the conditional gene knockout mice. The rat insulin promoter was cloned target the expression of Cre in pancreatic tissue. The Cre gene was modified by adding the nuclear localization signal and the sequence for initiation by eukaryotic ribosomes at 5' terminal of the Cre gene. Cre gene was linked to the intron of human growth factor gene. This construct was introduced into the mouse eggs using microinjection. Seven mice were identified as founders carrying the Cre gene by PCR. The results of RT-PCR showed that the transgenic mouse from one founder could transcribe the foreign gene in pancreas. The Southern blot analysis indicated that the Cre recombinase expressed in pancreas of the transgenic mouse was functional.
以组织特异性方式表达Cre重组酶的转基因小鼠是构建条件性基因敲除小鼠的有力工具。克隆大鼠胰岛素启动子以靶向Cre在胰腺组织中的表达。通过在Cre基因的5'末端添加核定位信号和真核核糖体起始序列对Cre基因进行修饰。Cre基因与人类生长因子基因的内含子相连。使用显微注射将该构建体导入小鼠卵中。通过PCR鉴定出七只小鼠为携带Cre基因的奠基者。RT-PCR结果表明,来自一位奠基者的转基因小鼠能够在胰腺中转录外源基因。Southern印迹分析表明,转基因小鼠胰腺中表达的Cre重组酶具有功能。