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 Previous Article

Eukaryotic Cell, May 2009, p. 800-804, Vol. 8, No. 5
1535-9778/09/$08.00+0     doi:10.1128/EC.00072-09
Copyright © 2009, American Society for Microbiology. All Rights Reserved.

Fungal Functional Genomics: Tunable Knockout-Knock-in Expression and Tagging Strategies{triangledown} ,{dagger}

Luis F. Larrondo, Hildur V. Colot, Christopher L. Baker, Jennifer J. Loros, and Jay C. Dunlap*

Department of Genetics, Dartmouth Medical School, Hanover, New Hampshire 03755

Received 4 March 2009/ Accepted 6 March 2009

Strategies for promoting high-efficiency homologous gene replacement have been developed and adopted for many filamentous fungal species. The next generation of analysis requires the ability to manipulate gene expression and to tag genes expressed from their endogenous loci. Here we present a suite of molecular tools that provide versatile solutions for fungal high-throughput functional genomics studies based on locus-specific modification of any target gene. Additionally, case studies illustrate caveats to presumed overexpression constructs. A tunable expression system and different tagging strategies can provide valuable phenotypic information for uncharacterized genes and facilitate the analysis of essential loci, an emerging problem in systematic deletion studies of haploid organisms.

* Corresponding author. Mailing address: Department of Genetics, Dartmouth Medical School, 7400 Remsen, Hanover, NH 03755. Phone: (603) 650-1108. Fax: (603) 650-1233. E-mail: Jay.C.Dunlap{at}Dartmouth.edu

{triangledown} Published ahead of print on 13 March 2009.

{dagger} Supplemental material for this article may be found at http://ec.asm.org/.

Eukaryotic Cell, May 2009, p. 800-804, Vol. 8, No. 5
1535-9778/09/$08.00+0     doi:10.1128/EC.00072-09
Copyright © 2009, American Society for Microbiology. All Rights Reserved.





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