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A universal TagModule collection for parallel genetic analysis of microorganisms

Systems-level analyses of non-model microorganisms are limited by the existence of numerous uncharacterized genes and a corresponding over-reliance on automated computational annotations. One solution to this challenge is to disrupt gene function using DNA tag technology, which has been highly succe...

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Autores principales: Oh, Julia, Fung, Eula, Price, Morgan N., Dehal, Paramvir S., Davis, Ronald W., Giaever, Guri, Nislow, Corey, Arkin, Adam P., Deutschbauer, Adam
Formato: Texto
Lenguaje:English
Publicado: Oxford University Press 2010
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2919733/
https://www.ncbi.nlm.nih.gov/pubmed/20494978
http://dx.doi.org/10.1093/nar/gkq419
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author Oh, Julia
Fung, Eula
Price, Morgan N.
Dehal, Paramvir S.
Davis, Ronald W.
Giaever, Guri
Nislow, Corey
Arkin, Adam P.
Deutschbauer, Adam
author_facet Oh, Julia
Fung, Eula
Price, Morgan N.
Dehal, Paramvir S.
Davis, Ronald W.
Giaever, Guri
Nislow, Corey
Arkin, Adam P.
Deutschbauer, Adam
author_sort Oh, Julia
collection PubMed
description Systems-level analyses of non-model microorganisms are limited by the existence of numerous uncharacterized genes and a corresponding over-reliance on automated computational annotations. One solution to this challenge is to disrupt gene function using DNA tag technology, which has been highly successful in parallelizing reverse genetics in Saccharomyces cerevisiae and has led to discoveries in gene function, genetic interactions and drug mechanism of action. To extend the yeast DNA tag methodology to a wide variety of microorganisms and applications, we have created a universal, sequence-verified TagModule collection. A hallmark of the 4280 TagModules is that they are cloned into a Gateway entry vector, thus facilitating rapid transfer to any compatible genetic system. Here, we describe the application of the TagModules to rapidly generate tagged mutants by transposon mutagenesis in the metal-reducing bacterium Shewanella oneidensis MR-1 and the pathogenic yeast Candida albicans. Our results demonstrate the optimal hybridization properties of the TagModule collection, the flexibility in applying the strategy to diverse microorganisms and the biological insights that can be gained from fitness profiling tagged mutant collections. The publicly available TagModule collection is a platform-independent resource for the functional genomics of a wide range of microbial systems in the post-genome era.
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spelling pubmed-29197332010-08-11 A universal TagModule collection for parallel genetic analysis of microorganisms Oh, Julia Fung, Eula Price, Morgan N. Dehal, Paramvir S. Davis, Ronald W. Giaever, Guri Nislow, Corey Arkin, Adam P. Deutschbauer, Adam Nucleic Acids Res Methods Online Systems-level analyses of non-model microorganisms are limited by the existence of numerous uncharacterized genes and a corresponding over-reliance on automated computational annotations. One solution to this challenge is to disrupt gene function using DNA tag technology, which has been highly successful in parallelizing reverse genetics in Saccharomyces cerevisiae and has led to discoveries in gene function, genetic interactions and drug mechanism of action. To extend the yeast DNA tag methodology to a wide variety of microorganisms and applications, we have created a universal, sequence-verified TagModule collection. A hallmark of the 4280 TagModules is that they are cloned into a Gateway entry vector, thus facilitating rapid transfer to any compatible genetic system. Here, we describe the application of the TagModules to rapidly generate tagged mutants by transposon mutagenesis in the metal-reducing bacterium Shewanella oneidensis MR-1 and the pathogenic yeast Candida albicans. Our results demonstrate the optimal hybridization properties of the TagModule collection, the flexibility in applying the strategy to diverse microorganisms and the biological insights that can be gained from fitness profiling tagged mutant collections. The publicly available TagModule collection is a platform-independent resource for the functional genomics of a wide range of microbial systems in the post-genome era. Oxford University Press 2010-08 2010-05-21 /pmc/articles/PMC2919733/ /pubmed/20494978 http://dx.doi.org/10.1093/nar/gkq419 Text en © The Author(s) 2010. Published by Oxford University Press. http://creativecommons.org/licenses/by-nc/2.5 This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/2.5), which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Methods Online
Oh, Julia
Fung, Eula
Price, Morgan N.
Dehal, Paramvir S.
Davis, Ronald W.
Giaever, Guri
Nislow, Corey
Arkin, Adam P.
Deutschbauer, Adam
A universal TagModule collection for parallel genetic analysis of microorganisms
title A universal TagModule collection for parallel genetic analysis of microorganisms
title_full A universal TagModule collection for parallel genetic analysis of microorganisms
title_fullStr A universal TagModule collection for parallel genetic analysis of microorganisms
title_full_unstemmed A universal TagModule collection for parallel genetic analysis of microorganisms
title_short A universal TagModule collection for parallel genetic analysis of microorganisms
title_sort universal tagmodule collection for parallel genetic analysis of microorganisms
topic Methods Online
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2919733/
https://www.ncbi.nlm.nih.gov/pubmed/20494978
http://dx.doi.org/10.1093/nar/gkq419
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