Quantitative analysis of triple-mutant genetic interactions
| العنوان: | Quantitative analysis of triple-mutant genetic interactions |
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| المؤلفون: | Michael Shales, Richard Alexander, James E. Haber, Hannes Braberg, Nevan J. Krogan, Qiuqin Wu, Jiewei Xu, Kathleen Franks-Skiba |
| المصدر: | Nature Protocols. 9:1867-1881 |
| بيانات النشر: | Springer Science and Business Media LLC, 2014. |
| سنة النشر: | 2014 |
| مصطلحات موضوعية: | Genetics, biology, High-throughput screening, Saccharomyces cerevisiae, Mutant, Gene regulatory network, Gene mutation, biology.organism_classification, Article, General Biochemistry, Genetics and Molecular Biology, Genes, Schizosaccharomyces, Epistasis, Gene Regulatory Networks, Gene, Gene Deletion, Genetic Association Studies |
| الوصف: | The quantitative analysis of genetic interactions between pairs of gene mutations has proven to be effective for characterizing cellular functions, but it can miss important interactions for functionally redundant genes. To address this limitation, we have developed an approach termed triple-mutant analysis (TMA). The procedure relies on a query strain that contains two deletions in a pair of redundant or otherwise related genes, which is crossed against a panel of candidate deletion strains to isolate triple mutants and measure their growth. A central feature of TMA is to interrogate mutants that are synthetically sick when two other genes are deleted but interact minimally with either single deletion. This approach has been valuable for discovering genes that restore critical functions when the principal actors are deleted. TMA has also uncovered double-mutant combinations that produce severe defects because a third protein becomes deregulated and acts in a deleterious fashion, and it has revealed functional differences between proteins presumed to act together. The protocol is optimized for Singer ROTOR pinning robots, takes 3 weeks to complete and measures interactions for up to 30 double mutants against a library of 1,536 single mutants. |
| تدمد: | 1750-2799 1754-2189 |
| URL الوصول: | https://explore.openaire.eu/search/publication?articleId=doi_dedup___::ac5c17aa47ca8ce7f7b5e86c31cdcd36 https://doi.org/10.1038/nprot.2014.127 |
| حقوق: | OPEN |
| رقم الأكسشن: | edsair.doi.dedup.....ac5c17aa47ca8ce7f7b5e86c31cdcd36 |
| قاعدة البيانات: | OpenAIRE |
| تدمد: | 17502799 17542189 |
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