| RRC ID |
35210
|
| 著者 |
Typas A, Nichols RJ, Siegele DA, Shales M, Collins SR, Lim B, Braberg H, Yamamoto N, Takeuchi R, Wanner BL, Mori H, Weissman JS, Krogan NJ, Gross CA.
|
| タイトル |
High-throughput, quantitative analyses of genetic interactions in E. coli.
|
| ジャーナル |
Nat Methods
|
| Abstract |
Large-scale genetic interaction studies provide the basis for defining gene function and pathway architecture. Recent advances in the ability to generate double mutants en masse in Saccharomyces cerevisiae have dramatically accelerated the acquisition of genetic interaction information and the biological inferences that follow. Here we describe a method based on F factor-driven conjugation, which allows for high-throughput generation of double mutants in Escherichia coli. This method, termed genetic interaction analysis technology for E. coli (GIANT-coli), permits us to systematically generate and array double-mutant cells on solid media in high-density arrays. We show that colony size provides a robust and quantitative output of cellular fitness and that GIANT-coli can recapitulate known synthetic interactions and identify previously unidentified negative (synthetic sickness or lethality) and positive (suppressive or epistatic) relationships. Finally, we describe a complementary strategy for genome-wide suppressor-mutant identification. Together, these methods permit rapid, large-scale genetic interaction studies in E. coli.
|
| 巻・号 |
5(9)
|
| ページ |
781-7
|
| 公開日 |
2008-9-1
|
| DOI |
10.1038/nmeth.1240
|
| PMID |
19160513
|
| PMC |
PMC2700713
|
| MeSH |
Conjugation, Genetic*
Escherichia coli / genetics*
Genome, Bacterial
Mutation*
|
| IF |
30.822
|
| 引用数 |
153
|
|
WOS 分野
|
BIOCHEMICAL RESEARCH METHODS
|
| リソース情報 |
| 原核生物(大腸菌) |
Keio collection
ASKA |
