遗传 ›› 2003, Vol. 25 ›› Issue (5): 628-632.

• 论文 • 上一篇    

Red重组系统及在微生物基因敲除中的应用

胡堃1;2;史兆兴1;赛道建2;黄留玉1 HU Kun1;2;SHI Zhao-Xing1;SAI Dao-Jian2;HUANG Liu-Yu1   

  1. 1.军事医学科学院生物工程研究所,北京 100071; 2.山东师范大学生命科学学院,济南 250014 1.Beijing Institute of Biotechnology,Beijing 100071,China; 2.College of Life Science of Shandong Normal University,Jinan 250014,China
  • 收稿日期:1900-01-01 出版日期:2003-10-10 发布日期:2003-10-10

The Red Recombination System and Its Application to Gene Knock-out in Microorganism

  • Received:1900-01-01 Published:2003-10-10 Online:2003-10-10

摘要: 在完成了对各种微生物基因组的测序以后,功能基因学的研究变得尤为重要。研究基因功能最直接的方法便是将待研究的基因失活。最初构建基因突变体是采用大肠杆菌的RecA系统,但是RecA重组系统操作复杂,重组效率低。最近建立了Red重组系统,该系统由3个蛋白组成:α蛋白(即λ核酸外切酶),β蛋白,Gam蛋白。应用Red系统进行基因敲除,可以直接利用线性打靶DNA,两侧同源臂长度在35~60 bp即可发生同源重组,且重组效率高。
Abstract:Since many DNA-sequencing projects of varied microorganisms have been completed,studies on their functional genomics become more important.Inactivation of an interesting gene is a direct method to characterize its function.Though the Esherichia coli RecA recombination system can be used to produce gene mutants,it needs a complex manipulation process.Furthermore,its efficiency is very low.Recently a Red recombination system was developed.This recombination system consists of three proteins:α protein(λ exonuclease),β protein and Gam protein.In this system,the linear targeting DNA which contains a selectable marker flanked with a homologous region as short as only 35~60 bp can be directly targeted for gene knock-out with a higher efficiency.

关键词: Red重组系统, 基因敲除, Red recombination system, gene knock-out, 抗药性基因
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