Identifying secreted proteins of Marssonina brunnea by degenerate PCR.
Identifieur interne : 003222 ( Main/Exploration ); précédent : 003221; suivant : 003223Identifying secreted proteins of Marssonina brunnea by degenerate PCR.
Auteurs : Qiang Cheng [République populaire de Chine] ; Youzhi Cao ; Cong Jiang ; Li'An Xu ; Mingxiu Wang ; Shougong Zhang ; Minren HuangSource :
- Proteomics [ 1615-9861 ] ; 2010.
Descripteurs français
- KwdFr :
- ADN complémentaire (génétique), Ascomycota (composition chimique), Ascomycota (métabolisme), Ascomycota (ultrastructure), Clonage moléculaire (MeSH), Données de séquences moléculaires (MeSH), Microscopie électronique à balayage (MeSH), Protéines fongiques (analyse), Protéines fongiques (génétique), RT-PCR (MeSH), Séquence d'acides aminés (MeSH), Séquence nucléotidique (MeSH).
- MESH :
- analyse : Protéines fongiques.
- composition chimique : Ascomycota.
- génétique : ADN complémentaire, Protéines fongiques.
- métabolisme : Ascomycota.
- ultrastructure : Ascomycota, Clonage moléculaire, Données de séquences moléculaires, Microscopie électronique à balayage, RT-PCR, Séquence d'acides aminés, Séquence nucléotidique.
English descriptors
- KwdEn :
- Amino Acid Sequence (MeSH), Ascomycota (chemistry), Ascomycota (metabolism), Ascomycota (ultrastructure), Base Sequence (MeSH), Cloning, Molecular (MeSH), DNA, Complementary (genetics), Fungal Proteins (analysis), Fungal Proteins (genetics), Microscopy, Electron, Scanning (MeSH), Molecular Sequence Data (MeSH), Reverse Transcriptase Polymerase Chain Reaction (MeSH).
- MESH :
- chemical , analysis : Fungal Proteins.
- chemical , genetics : DNA, Complementary, Fungal Proteins.
- chemistry : Ascomycota.
- metabolism : Ascomycota.
- ultrastructure : Ascomycota.
- Amino Acid Sequence, Base Sequence, Cloning, Molecular, Microscopy, Electron, Scanning, Molecular Sequence Data, Reverse Transcriptase Polymerase Chain Reaction.
Abstract
Marssonina brunnea is an important fungal pathogen of the Populus genus. To further our understanding of the pathogenesis of M. brunnea, we initiated a proteome-level study of the fungal secretome. Using de novo peptide sequencing by MS/MS, we obtained peptide sequences for 32 protein spots. Four proteins were identified by sequence homology to conserved proteins in public databases using MS-driven BLAST. To identify additional protein spots, we combined a degenerate PCR method, based on the Consensus-DEgenerate Hybrid Oligonucleotide Primer (CODEHOP) method, and a rapid amplification of cDNA ends method to clone the full-length cDNA fragments encoding the proteins identified in the gel. Using this method, we cloned the full-length cDNA fragments encoding 11 M. brunnea-specific proteins. This method provides an efficient approach to identification of species-specific proteins of non-sequenced organisms. Furthermore, we analyzed the expression patterns of these genes during infection. We found that most of the identified secreted proteins could be induced in artificial medium after hyphae entered poplar apoplast spaces. We propose that for the host-specialized M. brunnea, the elongation of hyphae has evolved closely with the secretion of apoplastic proteins.
DOI: 10.1002/pmic.200900844
PubMed: 20391531
Affiliations:
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sort="Wang, Mingxiu" uniqKey="Wang M" first="Mingxiu" last="Wang">Mingxiu Wang</name></author><author><name sortKey="Zhang, Shougong" sort="Zhang, Shougong" uniqKey="Zhang S" first="Shougong" last="Zhang">Shougong Zhang</name></author><author><name sortKey="Huang, Minren" sort="Huang, Minren" uniqKey="Huang M" first="Minren" last="Huang">Minren Huang</name></author></titleStmt><publicationStmt><idno type="wicri:source">PubMed</idno><date when="2010">2010</date><idno type="RBID">pubmed:20391531</idno><idno type="pmid">20391531</idno><idno type="doi">10.1002/pmic.200900844</idno><idno type="wicri:Area/Main/Corpus">003225</idno><idno type="wicri:explorRef" wicri:stream="Main" wicri:step="Corpus" wicri:corpus="PubMed">003225</idno><idno type="wicri:Area/Main/Curation">003225</idno><idno type="wicri:explorRef" wicri:stream="Main" wicri:step="Curation">003225</idno><idno type="wicri:Area/Main/Exploration">003225</idno></publicationStmt><sourceDesc><biblStruct><analytic><title xml:lang="en">Identifying secreted proteins of Marssonina brunnea by degenerate PCR.</title><author><name sortKey="Cheng, Qiang" sort="Cheng, Qiang" uniqKey="Cheng Q" first="Qiang" last="Cheng">Qiang Cheng</name><affiliation wicri:level="1"><nlm:affiliation>Nanjing Forestry University, Nanjing, People's Republic of China.</nlm:affiliation><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>Nanjing Forestry University, Nanjing</wicri:regionArea><wicri:noRegion>Nanjing</wicri:noRegion></affiliation></author><author><name sortKey="Cao, Youzhi" sort="Cao, Youzhi" uniqKey="Cao Y" first="Youzhi" last="Cao">Youzhi Cao</name></author><author><name sortKey="Jiang, Cong" sort="Jiang, Cong" uniqKey="Jiang C" first="Cong" last="Jiang">Cong Jiang</name></author><author><name sortKey="Xu, Li An" sort="Xu, Li An" uniqKey="Xu L" first="Li'An" last="Xu">Li'An Xu</name></author><author><name sortKey="Wang, Mingxiu" sort="Wang, Mingxiu" uniqKey="Wang M" first="Mingxiu" last="Wang">Mingxiu Wang</name></author><author><name sortKey="Zhang, Shougong" sort="Zhang, Shougong" uniqKey="Zhang S" first="Shougong" last="Zhang">Shougong Zhang</name></author><author><name sortKey="Huang, Minren" sort="Huang, Minren" uniqKey="Huang M" first="Minren" last="Huang">Minren Huang</name></author></analytic><series><title level="j">Proteomics</title><idno type="eISSN">1615-9861</idno><imprint><date when="2010" type="published">2010</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Amino Acid Sequence (MeSH)</term><term>Ascomycota (chemistry)</term><term>Ascomycota (metabolism)</term><term>Ascomycota (ultrastructure)</term><term>Base Sequence (MeSH)</term><term>Cloning, Molecular (MeSH)</term><term>DNA, Complementary (genetics)</term><term>Fungal Proteins (analysis)</term><term>Fungal Proteins (genetics)</term><term>Microscopy, Electron, Scanning (MeSH)</term><term>Molecular Sequence Data (MeSH)</term><term>Reverse Transcriptase Polymerase Chain Reaction (MeSH)</term></keywords><keywords scheme="KwdFr" xml:lang="fr"><term>ADN complémentaire (génétique)</term><term>Ascomycota (composition chimique)</term><term>Ascomycota (métabolisme)</term><term>Ascomycota (ultrastructure)</term><term>Clonage moléculaire (MeSH)</term><term>Données de séquences moléculaires (MeSH)</term><term>Microscopie électronique à balayage (MeSH)</term><term>Protéines fongiques (analyse)</term><term>Protéines fongiques (génétique)</term><term>RT-PCR (MeSH)</term><term>Séquence d'acides aminés (MeSH)</term><term>Séquence nucléotidique (MeSH)</term></keywords><keywords scheme="MESH" type="chemical" qualifier="analysis" xml:lang="en"><term>Fungal Proteins</term></keywords><keywords scheme="MESH" type="chemical" qualifier="genetics" xml:lang="en"><term>DNA, Complementary</term><term>Fungal Proteins</term></keywords><keywords scheme="MESH" qualifier="analyse" xml:lang="fr"><term>Protéines fongiques</term></keywords><keywords scheme="MESH" qualifier="chemistry" xml:lang="en"><term>Ascomycota</term></keywords><keywords scheme="MESH" qualifier="composition chimique" xml:lang="fr"><term>Ascomycota</term></keywords><keywords scheme="MESH" qualifier="génétique" xml:lang="fr"><term>ADN complémentaire</term><term>Protéines fongiques</term></keywords><keywords scheme="MESH" qualifier="metabolism" xml:lang="en"><term>Ascomycota</term></keywords><keywords scheme="MESH" qualifier="métabolisme" xml:lang="fr"><term>Ascomycota</term></keywords><keywords scheme="MESH" qualifier="ultrastructure" xml:lang="en"><term>Ascomycota</term></keywords><keywords scheme="MESH" xml:lang="en"><term>Amino Acid Sequence</term><term>Base Sequence</term><term>Cloning, Molecular</term><term>Microscopy, Electron, Scanning</term><term>Molecular Sequence Data</term><term>Reverse Transcriptase Polymerase Chain Reaction</term></keywords><keywords scheme="MESH" qualifier="ultrastructure" xml:lang="fr"><term>Ascomycota</term><term>Clonage moléculaire</term><term>Données de séquences moléculaires</term><term>Microscopie électronique à balayage</term><term>RT-PCR</term><term>Séquence d'acides aminés</term><term>Séquence nucléotidique</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">Marssonina brunnea is an important fungal pathogen of the Populus genus. To further our understanding of the pathogenesis of M. brunnea, we initiated a proteome-level study of the fungal secretome. Using de novo peptide sequencing by MS/MS, we obtained peptide sequences for 32 protein spots. Four proteins were identified by sequence homology to conserved proteins in public databases using MS-driven BLAST. To identify additional protein spots, we combined a degenerate PCR method, based on the Consensus-DEgenerate Hybrid Oligonucleotide Primer (CODEHOP) method, and a rapid amplification of cDNA ends method to clone the full-length cDNA fragments encoding the proteins identified in the gel. Using this method, we cloned the full-length cDNA fragments encoding 11 M. brunnea-specific proteins. This method provides an efficient approach to identification of species-specific proteins of non-sequenced organisms. Furthermore, we analyzed the expression patterns of these genes during infection. We found that most of the identified secreted proteins could be induced in artificial medium after hyphae entered poplar apoplast spaces. We propose that for the host-specialized M. brunnea, the elongation of hyphae has evolved closely with the secretion of apoplastic proteins.</div></front></TEI><pubmed><MedlineCitation Status="MEDLINE" Owner="NLM"><PMID Version="1">20391531</PMID><DateCompleted><Year>2010</Year><Month>09</Month><Day>24</Day></DateCompleted><DateRevised><Year>2010</Year><Month>08</Month><Day>23</Day></DateRevised><Article PubModel="Print"><Journal><ISSN IssnType="Electronic">1615-9861</ISSN><JournalIssue CitedMedium="Internet"><Volume>10</Volume><Issue>13</Issue><PubDate><Year>2010</Year><Month>Jul</Month></PubDate></JournalIssue><Title>Proteomics</Title><ISOAbbreviation>Proteomics</ISOAbbreviation></Journal><ArticleTitle>Identifying secreted proteins of Marssonina brunnea by degenerate PCR.</ArticleTitle><Pagination><MedlinePgn>2406-17</MedlinePgn></Pagination><ELocationID EIdType="doi" ValidYN="Y">10.1002/pmic.200900844</ELocationID><Abstract><AbstractText>Marssonina brunnea is an important fungal pathogen of the Populus genus. To further our understanding of the pathogenesis of M. brunnea, we initiated a proteome-level study of the fungal secretome. Using de novo peptide sequencing by MS/MS, we obtained peptide sequences for 32 protein spots. Four proteins were identified by sequence homology to conserved proteins in public databases using MS-driven BLAST. To identify additional protein spots, we combined a degenerate PCR method, based on the Consensus-DEgenerate Hybrid Oligonucleotide Primer (CODEHOP) method, and a rapid amplification of cDNA ends method to clone the full-length cDNA fragments encoding the proteins identified in the gel. Using this method, we cloned the full-length cDNA fragments encoding 11 M. brunnea-specific proteins. This method provides an efficient approach to identification of species-specific proteins of non-sequenced organisms. Furthermore, we analyzed the expression patterns of these genes during infection. We found that most of the identified secreted proteins could be induced in artificial medium after hyphae entered poplar apoplast spaces. We propose that for the host-specialized M. brunnea, the elongation of hyphae has evolved closely with the secretion of apoplastic proteins.</AbstractText></Abstract><AuthorList CompleteYN="Y"><Author ValidYN="Y"><LastName>Cheng</LastName><ForeName>Qiang</ForeName><Initials>Q</Initials><AffiliationInfo><Affiliation>Nanjing Forestry University, Nanjing, People's Republic of China.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Cao</LastName><ForeName>Youzhi</ForeName><Initials>Y</Initials></Author><Author ValidYN="Y"><LastName>Jiang</LastName><ForeName>Cong</ForeName><Initials>C</Initials></Author><Author ValidYN="Y"><LastName>Xu</LastName><ForeName>Li'an</ForeName><Initials>L</Initials></Author><Author ValidYN="Y"><LastName>Wang</LastName><ForeName>Mingxiu</ForeName><Initials>M</Initials></Author><Author ValidYN="Y"><LastName>Zhang</LastName><ForeName>Shougong</ForeName><Initials>S</Initials></Author><Author ValidYN="Y"><LastName>Huang</LastName><ForeName>Minren</ForeName><Initials>M</Initials></Author></AuthorList><Language>eng</Language><PublicationTypeList><PublicationType UI="D016428">Journal Article</PublicationType><PublicationType UI="D013485">Research Support, Non-U.S. Gov't</PublicationType></PublicationTypeList></Article><MedlineJournalInfo><Country>Germany</Country><MedlineTA>Proteomics</MedlineTA><NlmUniqueID>101092707</NlmUniqueID><ISSNLinking>1615-9853</ISSNLinking></MedlineJournalInfo><ChemicalList><Chemical><RegistryNumber>0</RegistryNumber><NameOfSubstance UI="D018076">DNA, Complementary</NameOfSubstance></Chemical><Chemical><RegistryNumber>0</RegistryNumber><NameOfSubstance UI="D005656">Fungal Proteins</NameOfSubstance></Chemical></ChemicalList><CitationSubset>IM</CitationSubset><MeshHeadingList><MeshHeading><DescriptorName UI="D000595" MajorTopicYN="N">Amino Acid Sequence</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D001203" MajorTopicYN="N">Ascomycota</DescriptorName><QualifierName UI="Q000737" MajorTopicYN="Y">chemistry</QualifierName><QualifierName UI="Q000378" MajorTopicYN="N">metabolism</QualifierName><QualifierName UI="Q000648" MajorTopicYN="N">ultrastructure</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D001483" MajorTopicYN="N">Base Sequence</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D003001" MajorTopicYN="N">Cloning, Molecular</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D018076" MajorTopicYN="N">DNA, Complementary</DescriptorName><QualifierName UI="Q000235" MajorTopicYN="N">genetics</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D005656" MajorTopicYN="N">Fungal Proteins</DescriptorName><QualifierName UI="Q000032" MajorTopicYN="Y">analysis</QualifierName><QualifierName UI="Q000235" MajorTopicYN="N">genetics</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D008855" MajorTopicYN="N">Microscopy, Electron, Scanning</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D008969" MajorTopicYN="N">Molecular Sequence Data</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D020133" MajorTopicYN="N">Reverse Transcriptase Polymerase Chain Reaction</DescriptorName></MeshHeading></MeshHeadingList></MedlineCitation><PubmedData><History><PubMedPubDate PubStatus="entrez"><Year>2010</Year><Month>4</Month><Day>15</Day><Hour>6</Hour><Minute>0</Minute></PubMedPubDate><PubMedPubDate PubStatus="pubmed"><Year>2010</Year><Month>4</Month><Day>15</Day><Hour>6</Hour><Minute>0</Minute></PubMedPubDate><PubMedPubDate PubStatus="medline"><Year>2010</Year><Month>9</Month><Day>25</Day><Hour>6</Hour><Minute>0</Minute></PubMedPubDate></History><PublicationStatus>ppublish</PublicationStatus><ArticleIdList><ArticleId IdType="pubmed">20391531</ArticleId><ArticleId IdType="doi">10.1002/pmic.200900844</ArticleId></ArticleIdList></PubmedData></pubmed><affiliations><list><country><li>République populaire de Chine</li></country></list><tree><noCountry><name sortKey="Cao, Youzhi" sort="Cao, Youzhi" uniqKey="Cao Y" first="Youzhi" last="Cao">Youzhi Cao</name><name sortKey="Huang, Minren" sort="Huang, Minren" uniqKey="Huang M" first="Minren" last="Huang">Minren Huang</name><name sortKey="Jiang, Cong" sort="Jiang, Cong" uniqKey="Jiang C" first="Cong" last="Jiang">Cong Jiang</name><name sortKey="Wang, Mingxiu" sort="Wang, Mingxiu" uniqKey="Wang M" first="Mingxiu" last="Wang">Mingxiu Wang</name><name sortKey="Xu, Li An" sort="Xu, Li An" uniqKey="Xu L" first="Li'An" last="Xu">Li'An Xu</name><name sortKey="Zhang, Shougong" sort="Zhang, Shougong" uniqKey="Zhang S" first="Shougong" last="Zhang">Shougong Zhang</name></noCountry><country name="République populaire de Chine"><noRegion><name sortKey="Cheng, Qiang" sort="Cheng, Qiang" uniqKey="Cheng Q" first="Qiang" last="Cheng">Qiang Cheng</name></noRegion></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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