J4 ›› 2011, Vol. 37 ›› Issue (2): 196-201.

• 基础研究 • 上一篇    下一篇



  1. 吉林大学畜牧兽医学院 动物胚胎工程吉林省重点实验室,吉林 长春130062
  • 收稿日期:2010-12-08 出版日期:2011-03-28 发布日期:2011-03-28
  • 通讯作者: 逄大欣(Tel:0431-87836175,E-mail:pdx@jlu.edu.cn);欧阳红生(Tel:0431-87836175,E-mail:ouyh@jlu.edu.cn) E-mail:pdx@jlu.edu.cn;ouyh@jlu.edu.cn
  • 作者简介:国家自然科学基金资助课题(30771582,130871841)
  • 基金资助:


Construction of Cre recombinase expression vector Cre-pCEP4 and its application in Cre/loxP system

ZHOU Yang,ZHU Jian-guo,TANG Xiao-chun,YAN Sen,SONG Na,ZHANG Ming-jun,LI Li,OU-YANG Hong-sheng,PANG Da-xin   

  1. Jilin Provincial Key Laboratory of Animal Embryo Engineering,Institute of Animal and Veterinary Medicine,Jilin University,Changchun 130062,China
  • Received:2010-12-08 Online:2011-03-28 Published:2011-03-28


目的:构建表达Cre重组酶的载体Cre-pCEP4,并验证其能够有效识别loxP位点,为人类疾病动物模型的建立提供依据。方法:构建重组载体Cre-pCEP4和pStop-eGFP,利用Fugene HD转染猪胚胎成纤维细胞(PEF)和MCF-7细胞系,利用荧光显微镜观察绿色荧光蛋白表达情况。结果:成功构建重组载体Cre-pCEP4和pStop-eGFP,将2个载体瞬时共转染PEF;经潮酶素B筛选出Cre重组酶稳定表达的MCF-7细胞系瞬时转染pStop-eGFP,在荧光显微镜下观察2种细胞均有绿色荧光蛋白的表达。而单独转染pStop-eGFP的MCF-7细胞系和PEF均未见绿色荧光蛋白的表达。结论:重组载体Cre-pCEP4在细胞内能够表达Cre重组酶,并且表达的Cre重组酶能够识别loxP位点,删除两同向loxP间的DNA片段。

关键词:  Cre-pCEP4;Cre/loxP重组系统;猪胚胎成纤维细胞;MCF-7细胞系


Abstract:Objective To construct the Cre rebombinase expression vector and identify its function of recognizing loxP sites and provide the basis for establishment of the animal models of human diseases. Methods The recombinant vectors Cre-pCEP4 and pStop-eGFP were constructed. The MCF-7 cells and porcine embryonic fibroblasts (PEF) were transfected by Fugene HD. The expression of eGFP was analyzed by fluorescence microscope. Results The recombinant vectors Cre-pCEP4 and pStop-eGFP were constructed successfully and they were co-transfected into PEF.The  MCF-7 cells with stable expression of Cre recombinase were transfected with pStop-eGFP after selection of hygromycin B. There were green fluorescent protein exrpression of eGFP analyzed by fluorescence microscope after transfected by the two vectors. No eGFP positive cell was detected in MCF-7 cells and PEF transfected with pStop-eGFP only. Conclusion The recombinant vector Cre-pCEP4 can express Cre recombinase which can recognize loxP sites and delete the DNA agment between two loxP sites in the same orientation.

Key words: Cre-pCEP4;Cre/loxP system;porcine embryonic fibroblasts;MCF-7 cell line


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