Document Detail


A universal TagModule collection for parallel genetic analysis of microorganisms.
MedLine Citation:
PMID:  20494978     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
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.
Authors:
Julia Oh; Eula Fung; Morgan N Price; Paramvir S Dehal; Ronald W Davis; Guri Giaever; Corey Nislow; Adam P Arkin; Adam Deutschbauer
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Publication Detail:
Type:  Journal Article; Research Support, N.I.H., Extramural; Research Support, U.S. Gov't, Non-P.H.S.     Date:  2010-05-21
Journal Detail:
Title:  Nucleic acids research     Volume:  38     ISSN:  1362-4962     ISO Abbreviation:  Nucleic Acids Res.     Publication Date:  2010 Aug 
Date Detail:
Created Date:  2010-08-12     Completed Date:  2010-09-23     Revised Date:  2010-09-30    
Medline Journal Info:
Nlm Unique ID:  0411011     Medline TA:  Nucleic Acids Res     Country:  England    
Other Details:
Languages:  eng     Pagination:  e146     Citation Subset:  IM    
Affiliation:
Department of Genetics, Stanford University, Palo Alto, CA 94305, USA.
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MeSH Terms
Descriptor/Qualifier:
Candida albicans / genetics*
DNA Transposable Elements
Mutagenesis, Insertional / methods*
Oligonucleotide Array Sequence Analysis
Sequence Tagged Sites
Shewanella / genetics*
Grant Support
ID/Acronym/Agency:
F32GM080968/GM/NIGMS NIH HHS; HG003317/HG/NHGRI NIH HHS; P01 GH000205/GH/COGH CDC HHS; R01 HG003317/HG/NHGRI NIH HHS; T32 HG00044/HG/NHGRI NIH HHS
Chemical
Reg. No./Substance:
0/DNA Transposable Elements
Comments/Corrections

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