Isolation and transformation of uracil auxotrophs of the lignin-degrading basidiomycete Phanerochaete chrysosporium.
Identifieur interne : 000E31 ( Main/Corpus ); précédent : 000E30; suivant : 000E32Isolation and transformation of uracil auxotrophs of the lignin-degrading basidiomycete Phanerochaete chrysosporium.
Auteurs : L. Akileswaran ; M. Alic ; E K Clark ; J L Hornick ; M H GoldSource :
- Current genetics [ 0172-8083 ] ; 1993.
English descriptors
- KwdEn :
- Ascomycota (genetics), Basidiomycota (genetics), Basidiomycota (metabolism), Drug Resistance, Microbial (genetics), Genes, Fungal (MeSH), Lignin (metabolism), Mutation (MeSH), Orotate Phosphoribosyltransferase (genetics), Orotic Acid (analogs & derivatives), Orotic Acid (pharmacology), Orotidine-5'-Phosphate Decarboxylase (genetics), Recombination, Genetic (MeSH), Transformation, Genetic (MeSH), Uracil (metabolism).
- MESH :
- chemical , analogs & derivatives : Orotic Acid.
- chemical , genetics : Orotate Phosphoribosyltransferase, Orotidine-5'-Phosphate Decarboxylase.
- chemical , metabolism : Lignin, Uracil.
- genetics : Ascomycota, Basidiomycota, Drug Resistance, Microbial.
- metabolism : Basidiomycota.
- chemical , pharmacology : Orotic Acid.
- Genes, Fungal, Mutation, Recombination, Genetic, Transformation, Genetic.
Abstract
Uracil auxotrophs of Phanerochaete chrysosporium were isolated using 5-fluoroorotate resistance as a selection scheme. The ura3 auxotrophs deficient in orotidylate decarboxylase and ura5 auxotrophs deficient in orotate phosphoribosyl transferase were characterized by enzyme assays and complementation tests. The ura5 auxotrophs were transformed to prototrophy with the ura5 gene from the ascomycete Podospora anserina. The ura3 auxotrophs were transformed to prototrophy with the ura3 gene from the basidiomycete Schizophyllum commune. The P. chrysosporium ura3 gene was isolated from a lambda EMBL3 genomic library using the S. commune ura3 gene as a probe. A 6.6-kb fragment incorporating the ura3 gene was subcloned into Bluescript SK+(pURA3.1) and used to transform P. chrysosporium ura3 auxotrophic strains. The pURA3.1 insert was mapped for restriction sites and the approximate location of the ura3 gene within the insert was determined. Double auxotrophic strains were transformed with either of two marker genes and the resulting single auxotrophic strains were crossed to demonstrate genetic recombination between two nuclei of identical genetic background.
DOI: 10.1007/BF00310898
PubMed: 8467534
Links to Exploration step
pubmed:8467534Le document en format XML
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<affiliation><nlm:affiliation>Department of Chemical and Biological Sciences, Oregon Graduate Institute of Science and Technology, Beaverton 97006-1999.</nlm:affiliation>
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<author><name sortKey="Alic, M" sort="Alic, M" uniqKey="Alic M" first="M" last="Alic">M. Alic</name>
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<author><name sortKey="Clark, E K" sort="Clark, E K" uniqKey="Clark E" first="E K" last="Clark">E K Clark</name>
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<author><name sortKey="Hornick, J L" sort="Hornick, J L" uniqKey="Hornick J" first="J L" last="Hornick">J L Hornick</name>
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<author><name sortKey="Gold, M H" sort="Gold, M H" uniqKey="Gold M" first="M H" last="Gold">M H Gold</name>
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<author><name sortKey="Alic, M" sort="Alic, M" uniqKey="Alic M" first="M" last="Alic">M. Alic</name>
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<author><name sortKey="Hornick, J L" sort="Hornick, J L" uniqKey="Hornick J" first="J L" last="Hornick">J L Hornick</name>
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<series><title level="j">Current genetics</title>
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<term>Basidiomycota (genetics)</term>
<term>Basidiomycota (metabolism)</term>
<term>Drug Resistance, Microbial (genetics)</term>
<term>Genes, Fungal (MeSH)</term>
<term>Lignin (metabolism)</term>
<term>Mutation (MeSH)</term>
<term>Orotate Phosphoribosyltransferase (genetics)</term>
<term>Orotic Acid (analogs & derivatives)</term>
<term>Orotic Acid (pharmacology)</term>
<term>Orotidine-5'-Phosphate Decarboxylase (genetics)</term>
<term>Recombination, Genetic (MeSH)</term>
<term>Transformation, Genetic (MeSH)</term>
<term>Uracil (metabolism)</term>
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<keywords scheme="MESH" type="chemical" qualifier="analogs & derivatives" xml:lang="en"><term>Orotic Acid</term>
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<keywords scheme="MESH" type="chemical" qualifier="genetics" xml:lang="en"><term>Orotate Phosphoribosyltransferase</term>
<term>Orotidine-5'-Phosphate Decarboxylase</term>
</keywords>
<keywords scheme="MESH" type="chemical" qualifier="metabolism" xml:lang="en"><term>Lignin</term>
<term>Uracil</term>
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<keywords scheme="MESH" qualifier="genetics" xml:lang="en"><term>Ascomycota</term>
<term>Basidiomycota</term>
<term>Drug Resistance, Microbial</term>
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<keywords scheme="MESH" qualifier="metabolism" xml:lang="en"><term>Basidiomycota</term>
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<keywords scheme="MESH" type="chemical" qualifier="pharmacology" xml:lang="en"><term>Orotic Acid</term>
</keywords>
<keywords scheme="MESH" xml:lang="en"><term>Genes, Fungal</term>
<term>Mutation</term>
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<term>Transformation, Genetic</term>
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<front><div type="abstract" xml:lang="en">Uracil auxotrophs of Phanerochaete chrysosporium were isolated using 5-fluoroorotate resistance as a selection scheme. The ura3 auxotrophs deficient in orotidylate decarboxylase and ura5 auxotrophs deficient in orotate phosphoribosyl transferase were characterized by enzyme assays and complementation tests. The ura5 auxotrophs were transformed to prototrophy with the ura5 gene from the ascomycete Podospora anserina. The ura3 auxotrophs were transformed to prototrophy with the ura3 gene from the basidiomycete Schizophyllum commune. The P. chrysosporium ura3 gene was isolated from a lambda EMBL3 genomic library using the S. commune ura3 gene as a probe. A 6.6-kb fragment incorporating the ura3 gene was subcloned into Bluescript SK+(pURA3.1) and used to transform P. chrysosporium ura3 auxotrophic strains. The pURA3.1 insert was mapped for restriction sites and the approximate location of the ura3 gene within the insert was determined. Double auxotrophic strains were transformed with either of two marker genes and the resulting single auxotrophic strains were crossed to demonstrate genetic recombination between two nuclei of identical genetic background.</div>
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<Issue>4</Issue>
<PubDate><Year>1993</Year>
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<Title>Current genetics</Title>
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<ArticleTitle>Isolation and transformation of uracil auxotrophs of the lignin-degrading basidiomycete Phanerochaete chrysosporium.</ArticleTitle>
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<Abstract><AbstractText>Uracil auxotrophs of Phanerochaete chrysosporium were isolated using 5-fluoroorotate resistance as a selection scheme. The ura3 auxotrophs deficient in orotidylate decarboxylase and ura5 auxotrophs deficient in orotate phosphoribosyl transferase were characterized by enzyme assays and complementation tests. The ura5 auxotrophs were transformed to prototrophy with the ura5 gene from the ascomycete Podospora anserina. The ura3 auxotrophs were transformed to prototrophy with the ura3 gene from the basidiomycete Schizophyllum commune. The P. chrysosporium ura3 gene was isolated from a lambda EMBL3 genomic library using the S. commune ura3 gene as a probe. A 6.6-kb fragment incorporating the ura3 gene was subcloned into Bluescript SK+(pURA3.1) and used to transform P. chrysosporium ura3 auxotrophic strains. The pURA3.1 insert was mapped for restriction sites and the approximate location of the ura3 gene within the insert was determined. Double auxotrophic strains were transformed with either of two marker genes and the resulting single auxotrophic strains were crossed to demonstrate genetic recombination between two nuclei of identical genetic background.</AbstractText>
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