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Article: Gene discovery and gene function assignment in filamentous fungi

TitleGene discovery and gene function assignment in filamentous fungi
Authors
Issue Date2001
PublisherNational Academy of Sciences. The Journal's web site is located at http://www.pnas.org
Citation
Proceedings Of The National Academy Of Sciences Of The United States Of America, 2001, v. 98 n. 9, p. 5110-5115 How to Cite?
AbstractFilamentous fungi are a large group of diverse and economically important microorganisms. Large-scale gene disruption strategies developed in budding yeast are not applicable to these organisms because of their larger genomes and lower rate of targeted integration (Tl) during transformation. We developed transposon-arrayed gene knockouts (TAGKO) to discover genes and simultaneously create gene disruption cassettes for subsequent transformation and mutant analysis. Transposons carrying a bacterial and fungal drug resistance marker are used to mutagenize individual cosmids or entire libraries in vitro. Cosmids are annotated by DNA sequence analysis at the transposon insertion sites, and cosmid inserts are liberated to direct insertional mutagenesis events in the genome. Based on saturation analysis of a cosmid insert and insertions in a fungal cosmid library, we show that TAGKO can be used to rapidly identify and mutate genes. We further show that insertions can create alterations in gene expression, and we have used this approach to investigate an amino acid oxidation pathway in two important fungal phytopathogens.
Persistent Identifierhttp://hdl.handle.net/10722/178760
ISSN
2015 Impact Factor: 9.423
2015 SCImago Journal Rankings: 6.883
ISI Accession Number ID
References

 

DC FieldValueLanguage
dc.contributor.authorHamer, Len_US
dc.contributor.authorAdachi, Ken_US
dc.contributor.authorMontenegroChamorro, MVen_US
dc.contributor.authorTanzer, MMen_US
dc.contributor.authorMahanty, SKen_US
dc.contributor.authorLo, Cen_US
dc.contributor.authorTarpey, RWen_US
dc.contributor.authorSkalchunes, ARen_US
dc.contributor.authorHeiniger, RWen_US
dc.contributor.authorFrank, SAen_US
dc.contributor.authorDarveaux, BAen_US
dc.contributor.authorLampe, DJen_US
dc.contributor.authorSlater, TMen_US
dc.contributor.authorRamamurthy, Len_US
dc.contributor.authorDezwaan, TMen_US
dc.contributor.authorNelson, GHen_US
dc.contributor.authorShuster, JRen_US
dc.contributor.authorWoessner, Jen_US
dc.contributor.authorHamer, JEen_US
dc.date.accessioned2012-12-19T09:49:35Z-
dc.date.available2012-12-19T09:49:35Z-
dc.date.issued2001en_US
dc.identifier.citationProceedings Of The National Academy Of Sciences Of The United States Of America, 2001, v. 98 n. 9, p. 5110-5115en_US
dc.identifier.issn0027-8424en_US
dc.identifier.urihttp://hdl.handle.net/10722/178760-
dc.description.abstractFilamentous fungi are a large group of diverse and economically important microorganisms. Large-scale gene disruption strategies developed in budding yeast are not applicable to these organisms because of their larger genomes and lower rate of targeted integration (Tl) during transformation. We developed transposon-arrayed gene knockouts (TAGKO) to discover genes and simultaneously create gene disruption cassettes for subsequent transformation and mutant analysis. Transposons carrying a bacterial and fungal drug resistance marker are used to mutagenize individual cosmids or entire libraries in vitro. Cosmids are annotated by DNA sequence analysis at the transposon insertion sites, and cosmid inserts are liberated to direct insertional mutagenesis events in the genome. Based on saturation analysis of a cosmid insert and insertions in a fungal cosmid library, we show that TAGKO can be used to rapidly identify and mutate genes. We further show that insertions can create alterations in gene expression, and we have used this approach to investigate an amino acid oxidation pathway in two important fungal phytopathogens.en_US
dc.languageengen_US
dc.publisherNational Academy of Sciences. The Journal's web site is located at http://www.pnas.orgen_US
dc.relation.ispartofProceedings of the National Academy of Sciences of the United States of Americaen_US
dc.subject.meshAllelesen_US
dc.subject.meshAscomycota - Geneticsen_US
dc.subject.meshCloning, Molecularen_US
dc.subject.meshCosmids - Geneticsen_US
dc.subject.meshCrops, Agricultural - Microbiologyen_US
dc.subject.meshDna Transposable Elements - Geneticsen_US
dc.subject.meshFungal Proteins - Genetics - Physiologyen_US
dc.subject.meshGene Deletionen_US
dc.subject.meshGene Expression Regulation, Fungalen_US
dc.subject.meshGenes, Fungal - Genetics - Physiologyen_US
dc.subject.meshGenomic Libraryen_US
dc.subject.meshMadurella - Geneticsen_US
dc.subject.meshMutagenesis, Insertional - Geneticsen_US
dc.subject.meshMutagenesis, Site-Directed - Geneticsen_US
dc.subject.meshPhenotypeen_US
dc.subject.meshReproducibility Of Resultsen_US
dc.subject.meshSequence Analysis, Dnaen_US
dc.subject.meshTransformation, Geneticen_US
dc.titleGene discovery and gene function assignment in filamentous fungien_US
dc.typeArticleen_US
dc.identifier.emailLo, C: clivelo@hkucc.hku.hken_US
dc.identifier.authorityLo, C=rp00751en_US
dc.description.naturelink_to_subscribed_fulltexten_US
dc.identifier.doi10.1073/pnas.091094198en_US
dc.identifier.pmid11296265-
dc.identifier.scopuseid_2-s2.0-0035942235en_US
dc.relation.referenceshttp://www.scopus.com/mlt/select.url?eid=2-s2.0-0035942235&selection=ref&src=s&origin=recordpageen_US
dc.identifier.volume98en_US
dc.identifier.issue9en_US
dc.identifier.spage5110en_US
dc.identifier.epage5115en_US
dc.identifier.isiWOS:000168311500057-
dc.publisher.placeUnited Statesen_US
dc.identifier.scopusauthoridHamer, L=6603685512en_US
dc.identifier.scopusauthoridAdachi, K=7401602553en_US
dc.identifier.scopusauthoridMontenegroChamorro, MV=7801549552en_US
dc.identifier.scopusauthoridTanzer, MM=7006052511en_US
dc.identifier.scopusauthoridMahanty, SK=7006558473en_US
dc.identifier.scopusauthoridLo, C=15737175700en_US
dc.identifier.scopusauthoridTarpey, RW=6506649352en_US
dc.identifier.scopusauthoridSkalchunes, AR=6506371230en_US
dc.identifier.scopusauthoridHeiniger, RW=6603138737en_US
dc.identifier.scopusauthoridFrank, SA=7202570325en_US
dc.identifier.scopusauthoridDarveaux, BA=8722585600en_US
dc.identifier.scopusauthoridLampe, DJ=7005684863en_US
dc.identifier.scopusauthoridSlater, TM=7101827995en_US
dc.identifier.scopusauthoridRamamurthy, L=7004041098en_US
dc.identifier.scopusauthoridDeZwaan, TM=6506211019en_US
dc.identifier.scopusauthoridNelson, GH=7402779849en_US
dc.identifier.scopusauthoridShuster, JR=7201487088en_US
dc.identifier.scopusauthoridWoessner, J=7004957337en_US
dc.identifier.scopusauthoridHamer, JE=7102288376en_US

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