CAJ | 학술논문

组织特异性表达Cre重组酶的转基因小鼠是进行组织特异性基因剔除研究的重要工具.为了建立胰腺组织特异性Cre转基因小鼠,我们通过PCR克隆了大鼠胰岛素基因启动子,并用它指导Cre基因在胰岛细胞中的特异性表达.在Cre重组酶基因5'端添加了真核核糖体结合序列和核定位序列以使Cre重组酶能穿越核膜在细胞核中发挥功能;同时,在Cre基因3'端添加了含内含子的3'端人生长激素基因.表达载体经显微注射导入小鼠受精卵以建立转基因小鼠.PCR检测显示共获得7只Cre整合阳性的转基因首建者小鼠;RT-PCR结果表明其中1只首建者小鼠的子代鼠在胰腺中转录了外源基因,进一步的Southem杂交结果表明,该转基因小鼠能够在胰腺中表达有功能的Cre重组酶.
조직특이성표체Cre중조매적전기인소서시진행조직특이성기인척제연구적중요공구.위료건립이선조직특이성Cre전기인소서,아문통과PCR극륭료대서이도소기인계동자,병용타지도Cre기인재이도세포중적특이성표체.재Cre중조매기인5'단첨가료진핵핵당체결합서렬화핵정위서렬이사Cre중조매능천월핵막재세포핵중발휘공능;동시,재Cre기인3'단첨가료함내함자적3'단인생장격소기인.표체재체경현미주사도입소서수정란이건립전기인소서.PCR검측현시공획득7지Cre정합양성적전기인수건자소서;RT-PCR결과표명기중1지수건자소서적자대서재이선중전록료외원기인,진일보적Southem잡교결과표명,해전기인소서능구재이선중표체유공능적Cre중조매.
The trantional 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 Crc recombinase expressed in pancreas of the transgenic mouse was functional.