Recombineering BAC transgenes for protein tagging

被引：22

作者：

Ciotta, Giovanni

Hofemeister, Helmut

Maresca, Marcello

Fu, Jun

Sarov, Mihail ^{[2
]}

Anastassiadis, Konstantinos ^{[1
]}

Stewart, A. Francis ^{[1
]}

机构：

[1] Tech Univ Dresden, BioInnovat Zentrum, Ctr Regenerat Therapy Dresden, D-01307 Dresden, Germany

[2] Max Planck Inst Cell Biol & Genet, D-01307 Dresden, Germany

来源：

METHODS | 2011年 / 53卷 / 02期

关键词：

BACTERIAL ARTIFICIAL CHROMOSOMES; SACCHAROMYCES-CEREVISIAE; ESCHERICHIA-COLI; HOMOLOGOUS RECOMBINATION; FUNCTIONAL GENOMICS; MOUSE GENOME; DNA; PURIFICATION; EXPLORATION; EFFICIENT;

D O I：

10.1016/j.ymeth.2010.09.003

中图分类号：

Q5 [生物化学];

学科分类号：

070307 [化学生物学];

摘要：

Protein tagging offers many advantages for proteomic and regulomic research, particularly due to the use of generic and highly sensitive methods that can be applied with reasonable throughput. Ideally, protein tagging is equivalent to having a high affinity antibody for every chosen protein. However, these advantages are compromised if the tagged protein is overexpressed, which is usually the case from cDNA expression vectors. BAC (bacterial artificial chromosome) transgenes present a way to express a chosen protein at physiological levels with all regulatory elements in their native configurations, including cell cycle, alternative splicing and microRNA regulation. Recombineering has become the method of choice for modifying large constructs like BACs. Here, we present a method for protein tagging by recombineering BACs, transfecting cells and evaluating tagged protein expression. (C) 2011 Published by Elsevier Inc.

引用

页码：113 / 119

页数：7

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