A homokaryotic derivative of a Phanerochaete chrysosporium strain and its use in genomic analysis of repetitive elements.
Identifieur interne : 000B01 ( Main/Corpus ); précédent : 000B00; suivant : 000B02A homokaryotic derivative of a Phanerochaete chrysosporium strain and its use in genomic analysis of repetitive elements.
Auteurs : P. Stewart ; J. Gaskell ; D. CullenSource :
- Applied and environmental microbiology [ 0099-2240 ] ; 2000.
English descriptors
- KwdEn :
- Chromosome Mapping (MeSH), Culture Media (MeSH), Genetic Linkage (MeSH), Karyotyping (MeSH), Molecular Sequence Data (MeSH), Peroxidases (genetics), Peroxidases (metabolism), Phanerochaete (enzymology), Phanerochaete (genetics), Phanerochaete (growth & development), Protoplasts (MeSH), Repetitive Sequences, Nucleic Acid (genetics), Reverse Transcriptase Polymerase Chain Reaction (MeSH), Sequence Analysis, DNA (MeSH), Transcription, Genetic (MeSH).
- MESH :
- chemical , genetics : Peroxidases.
- chemical , metabolism : Peroxidases.
- chemical : Culture Media.
- enzymology : Phanerochaete.
- genetics : Phanerochaete, Repetitive Sequences, Nucleic Acid.
- growth & development : Phanerochaete.
- Chromosome Mapping, Genetic Linkage, Karyotyping, Molecular Sequence Data, Protoplasts, Reverse Transcriptase Polymerase Chain Reaction, Sequence Analysis, DNA, Transcription, Genetic.
Abstract
Analysis of complex gene families in the lignin-degrading basidiomycete Phanerochaete chrysosporium has been hampered by the dikaryotic nuclear condition. To facilitate genetic investigations in P. chrysosporium strain BKM-F-1767, we isolated a homokaryon from regenerated protoplasts. The nuclear condition was established by PCR amplification of five unlinked genes followed by probing with allele-specific oligonucleotides. Under standard nitrogen-limited culture conditions, lignin peroxidase, manganese peroxidase, and glyoxal oxidase activities of the homokaryon were equivalent to those of the parental dikaryon. We used the homokaryon to determine the genomic organization and to assess transcriptional effects of a family of repetitive elements. Previous studies had identified an insertional mutation, Pce1, within lignin peroxidase allele lipI2. The element resembled nonautonomous class II transposons and was present in multiple copies in strain BKM-F-1767. In the present study, three additional copies of the Pce1-like element were cloned and sequenced. The distribution of elements was nonrandom; all localized to the same 3.7-Mb chromosome, as assessed by segregation analysis and Southern blot analysis of the homokaryon. Reverse transcription-PCR (RT-PCR) showed that Pce1 was not spliced from the lipI2 transcript in either the homokaryon or the parental dikaryon. However, both strains had equivalent lignin peroxidase activity, suggesting that some lip genes may be redundant.
DOI: 10.1128/aem.66.4.1629-1633.2000
PubMed: 10742252
PubMed Central: PMC92033
Links to Exploration step
pubmed:10742252Le document en format XML
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Cullen</name></author></titleStmt><publicationStmt><idno type="wicri:source">PubMed</idno><date when="2000">2000</date><idno type="RBID">pubmed:10742252</idno><idno type="pmid">10742252</idno><idno type="pmc">PMC92033</idno><idno type="doi">10.1128/aem.66.4.1629-1633.2000</idno><idno type="wicri:Area/Main/Corpus">000B01</idno><idno type="wicri:explorRef" wicri:stream="Main" wicri:step="Corpus" wicri:corpus="PubMed">000B01</idno></publicationStmt><sourceDesc><biblStruct><analytic><title xml:lang="en">A homokaryotic derivative of a Phanerochaete chrysosporium strain and its use in genomic analysis of repetitive elements.</title><author><name sortKey="Stewart, P" sort="Stewart, P" uniqKey="Stewart P" first="P" last="Stewart">P. Stewart</name><affiliation><nlm:affiliation>Department of Bacteriology, University of Wisconsin-Madison, Madison, Wisconsin 53706, USA.</nlm:affiliation></affiliation></author><author><name sortKey="Gaskell, J" sort="Gaskell, J" uniqKey="Gaskell J" first="J" last="Gaskell">J. Gaskell</name></author><author><name sortKey="Cullen, D" sort="Cullen, D" uniqKey="Cullen D" first="D" last="Cullen">D. Cullen</name></author></analytic><series><title level="j">Applied and environmental microbiology</title><idno type="ISSN">0099-2240</idno><imprint><date when="2000" type="published">2000</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Chromosome Mapping (MeSH)</term><term>Culture Media (MeSH)</term><term>Genetic Linkage (MeSH)</term><term>Karyotyping (MeSH)</term><term>Molecular Sequence Data (MeSH)</term><term>Peroxidases (genetics)</term><term>Peroxidases (metabolism)</term><term>Phanerochaete (enzymology)</term><term>Phanerochaete (genetics)</term><term>Phanerochaete (growth & development)</term><term>Protoplasts (MeSH)</term><term>Repetitive Sequences, Nucleic Acid (genetics)</term><term>Reverse Transcriptase Polymerase Chain Reaction (MeSH)</term><term>Sequence Analysis, DNA (MeSH)</term><term>Transcription, Genetic (MeSH)</term></keywords><keywords scheme="MESH" type="chemical" qualifier="genetics" xml:lang="en"><term>Peroxidases</term></keywords><keywords scheme="MESH" type="chemical" qualifier="metabolism" xml:lang="en"><term>Peroxidases</term></keywords><keywords scheme="MESH" type="chemical" xml:lang="en"><term>Culture Media</term></keywords><keywords scheme="MESH" qualifier="enzymology" xml:lang="en"><term>Phanerochaete</term></keywords><keywords scheme="MESH" qualifier="genetics" xml:lang="en"><term>Phanerochaete</term><term>Repetitive Sequences, Nucleic Acid</term></keywords><keywords scheme="MESH" qualifier="growth & development" xml:lang="en"><term>Phanerochaete</term></keywords><keywords scheme="MESH" xml:lang="en"><term>Chromosome Mapping</term><term>Genetic Linkage</term><term>Karyotyping</term><term>Molecular Sequence Data</term><term>Protoplasts</term><term>Reverse Transcriptase Polymerase Chain Reaction</term><term>Sequence Analysis, DNA</term><term>Transcription, Genetic</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">Analysis of complex gene families in the lignin-degrading basidiomycete Phanerochaete chrysosporium has been hampered by the dikaryotic nuclear condition. To facilitate genetic investigations in P. chrysosporium strain BKM-F-1767, we isolated a homokaryon from regenerated protoplasts. The nuclear condition was established by PCR amplification of five unlinked genes followed by probing with allele-specific oligonucleotides. Under standard nitrogen-limited culture conditions, lignin peroxidase, manganese peroxidase, and glyoxal oxidase activities of the homokaryon were equivalent to those of the parental dikaryon. We used the homokaryon to determine the genomic organization and to assess transcriptional effects of a family of repetitive elements. Previous studies had identified an insertional mutation, Pce1, within lignin peroxidase allele lipI2. The element resembled nonautonomous class II transposons and was present in multiple copies in strain BKM-F-1767. In the present study, three additional copies of the Pce1-like element were cloned and sequenced. The distribution of elements was nonrandom; all localized to the same 3.7-Mb chromosome, as assessed by segregation analysis and Southern blot analysis of the homokaryon. Reverse transcription-PCR (RT-PCR) showed that Pce1 was not spliced from the lipI2 transcript in either the homokaryon or the parental dikaryon. However, both strains had equivalent lignin peroxidase activity, suggesting that some lip genes may be redundant.</div></front></TEI><pubmed><MedlineCitation Status="MEDLINE" Owner="NLM"><PMID Version="1">10742252</PMID><DateCompleted><Year>2000</Year><Month>05</Month><Day>11</Day></DateCompleted><DateRevised><Year>2019</Year><Month>05</Month><Day>08</Day></DateRevised><Article PubModel="Print"><Journal><ISSN IssnType="Print">0099-2240</ISSN><JournalIssue CitedMedium="Print"><Volume>66</Volume><Issue>4</Issue><PubDate><Year>2000</Year><Month>Apr</Month></PubDate></JournalIssue><Title>Applied and environmental microbiology</Title><ISOAbbreviation>Appl Environ Microbiol</ISOAbbreviation></Journal><ArticleTitle>A homokaryotic derivative of a Phanerochaete chrysosporium strain and its use in genomic analysis of repetitive elements.</ArticleTitle><Pagination><MedlinePgn>1629-33</MedlinePgn></Pagination><Abstract><AbstractText>Analysis of complex gene families in the lignin-degrading basidiomycete Phanerochaete chrysosporium has been hampered by the dikaryotic nuclear condition. To facilitate genetic investigations in P. chrysosporium strain BKM-F-1767, we isolated a homokaryon from regenerated protoplasts. The nuclear condition was established by PCR amplification of five unlinked genes followed by probing with allele-specific oligonucleotides. Under standard nitrogen-limited culture conditions, lignin peroxidase, manganese peroxidase, and glyoxal oxidase activities of the homokaryon were equivalent to those of the parental dikaryon. We used the homokaryon to determine the genomic organization and to assess transcriptional effects of a family of repetitive elements. Previous studies had identified an insertional mutation, Pce1, within lignin peroxidase allele lipI2. The element resembled nonautonomous class II transposons and was present in multiple copies in strain BKM-F-1767. In the present study, three additional copies of the Pce1-like element were cloned and sequenced. The distribution of elements was nonrandom; all localized to the same 3.7-Mb chromosome, as assessed by segregation analysis and Southern blot analysis of the homokaryon. Reverse transcription-PCR (RT-PCR) showed that Pce1 was not spliced from the lipI2 transcript in either the homokaryon or the parental dikaryon. However, both strains had equivalent lignin peroxidase activity, suggesting that some lip genes may be redundant.</AbstractText></Abstract><AuthorList CompleteYN="Y"><Author ValidYN="Y"><LastName>Stewart</LastName><ForeName>P</ForeName><Initials>P</Initials><AffiliationInfo><Affiliation>Department of Bacteriology, University of Wisconsin-Madison, Madison, Wisconsin 53706, USA.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Gaskell</LastName><ForeName>J</ForeName><Initials>J</Initials></Author><Author ValidYN="Y"><LastName>Cullen</LastName><ForeName>D</ForeName><Initials>D</Initials></Author></AuthorList><Language>eng</Language><DataBankList CompleteYN="Y"><DataBank><DataBankName>GENBANK</DataBankName><AccessionNumberList><AccessionNumber>AF134289</AccessionNumber><AccessionNumber>AF134290</AccessionNumber><AccessionNumber>AF134291</AccessionNumber></AccessionNumberList></DataBank></DataBankList><PublicationTypeList><PublicationType UI="D016428">Journal 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