A RECOMBINEERING PIPELINE TO MAKE CONDITIONAL TARGETING CONSTRUCTS

被引：70

作者：

Fu, Jun ^{[1
]}

Teucher, Madeleine ^{[1
]}

Anastassiadis, Konstantinos ^{[2
]}

Skarnes, William ^{[3
]}

Stewart, A. Francis ^{[1
]}

机构：

[1] Tech Univ Dresden, Genom BioInnovationsZentrum, Dresden, Germany

[2] Tech Univ Dresden, Ctr Regenerat Therapy, Dresden, Germany

[3] Wellcome Trust Sanger Inst, Cambridge, England

来源：

METHODS IN ENZYMOLOGY, VOL 477: GUIDE TO TECHNIQUES IN MOUSE DEVELOPMENT, PART B: MOUSE MOLECULAR GENETICS, SECOND EDITION | 2010年 / 477卷

关键词：

BACTERIAL ARTIFICIAL CHROMOSOMES; EMBRYONIC STEM-CELLS; ESCHERICHIA-COLI; HOMOLOGOUS RECOMBINATION; MOUSE GENOME; IN-VIVO; FUNCTIONAL GENOMICS; GENE; MICE; CRE;

D O I：

10.1016/S0076-6879(10)77008-7

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

Here we describe an optimized recombineering protocol for the creation of conditional alleles. Recombineering is a DNA engineering methodology based on homologous recombination in Escherichia coli. It presents a number of advantages over conventional engineering methods, including the ability to place target sites for site-specific recombinases at exactly chosen positions. Consequently, complicated designs, such as conditional alleles, can be rapidly achieved. We have applied these advantages to establish a pipeline method for the fluent generation of conditional alleles which employs Cre, Flp, and Dre recombinase target sites.

引用

页码：125 / 144

页数：20

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