遗传 ›› 2007, Vol. 29 ›› Issue (5): 570-570―574.doi: 10.1360/yc-007-0570

• 研究报告 • 上一篇    下一篇

利用Red同源重组系统进行牛β酪蛋白基因敲除

薛可1,2; 李峰3; 罗光彬1; 黄玮玮2,4; 陈学进2   

  1. 1. 沈阳农业大学畜牧兽医学院, 沈阳 110161;
    2. 上海交通大学附属新华医院发育生物学研究中心, 上海 200092;
    3. 中国科学院上海生命科学研究院生物化学与细胞生物学研究所, 上海 200031;
    4. 华南农业大学动物科学学院, 广州 510642

  • 收稿日期:2006-10-30 修回日期:2006-11-30 出版日期:2007-05-08 发布日期:2007-05-08
  • 通讯作者: 陈学进

Knock out of bovine beta casein gene by homologous recombination

XUE Ke1,2; LI Feng3; LUO Guang-Bin1; HUANG Wei-Wei2,4; CHEN Xue-Jin2   

  1. 1. College of Animal Science and Vet Medicine of Shenyang Agricultural University, Shenyang 110161, China;
    2. Shanghai Jiao Tong University Xinhua Hospital Development Biology Research Center, Shanghai 200092, China;
    3. Institutes of Biochemistry and Cell Biology, SIBS, CAS, Shanghai 200031,China;
    4. College of Animal Science of Huanan Agricultural University, Guangzhou 510642, China
  • Received:2006-10-30 Revised:2006-11-30 Online:2007-05-08 Published:2007-05-08
  • Contact: CHEN Xue-Jin

摘要:

利用EL350基因工程菌进行同源重组, 成功进行基因敲除已有报道, 但利用该系统进行乳腺生物反应器质粒构建的研究却未见报道。实验采用含有完整的牛b 酪蛋白基因的CSN2质粒作为基因打靶的载体, 设计不同的同源臂, 成功地敲除了b 酪蛋白基因的编码区。并且同时利用同源重组技术对敲除不同大小的DNA片段的效率进行了研究。为进一步利用CSN2质粒两端的调控序列, 插入新的基因, 研究其表达功能, 或者进行乳腺生物反应器的研究奠定了基础。

关键词: 同源重组, 牛b酪蛋白基因, 基因敲除, Cre-loxP重组系统

Abstract:

It has been reported that homologous recombination with Red system has been successfully used for knock-out. We try to work on the construction of the expression vector of Mammary Gland with Red system. This study takes CSN2 as a vector for gene target, which contains the complete bovine beta casein gene. Different homologous arms were designed and the CDS region of the beta casein gene was successfully knocked out. The efficiency was also explored for knocking out different DNA fragment. Based on the study, it is very convenient for making a deep research of the foreign gene expression under the regulation of CSN2 flanking region.