A native promoter and inclusion of an intron is necessary for efficient expression of GFP or mRFP in Armillaria mellea.
Identifieur interne : 000230 ( Main/Exploration ); précédent : 000229; suivant : 000231A native promoter and inclusion of an intron is necessary for efficient expression of GFP or mRFP in Armillaria mellea.
Auteurs : Kathryn L. Ford [Royaume-Uni] ; Kendra Baumgartner [États-Unis] ; Béatrice Henricot [Royaume-Uni] ; Andy M. Bailey [Royaume-Uni] ; Gary D. Foster [Royaume-Uni]Source :
- Scientific reports [ 2045-2322 ] ; 2016.
Descripteurs français
- KwdFr :
- MESH :
English descriptors
- KwdEn :
- MESH :
- chemical , genetics : Green Fluorescent Proteins, Nuclear Proteins.
- genetics : Agaricus, Armillaria, Inclusion Bodies, Introns, Mycelium, Phanerochaete, Promoter Regions, Genetic.
Abstract
Armillaria mellea is a significant pathogen that causes Armillaria root disease on numerous hosts in forests, gardens and agricultural environments worldwide. Using a yeast-adapted pCAMBIA0380 Agrobacterium vector, we have constructed a series of vectors for transformation of A. mellea, assembled using yeast-based recombination methods. These have been designed to allow easy exchange of promoters and inclusion of introns. The vectors were first tested by transformation into basidiomycete Clitopilus passeckerianus to ascertain vector functionality then used to transform A. mellea. We show that heterologous promoters from the basidiomycetes Agaricus bisporus and Phanerochaete chrysosporium that were used successfully to control the hygromycin resistance cassette were not able to support expression of mRFP or GFP in A. mellea. The endogenous A. mellea gpd promoter delivered efficient expression, and we show that inclusion of an intron was also required for transgene expression. GFP and mRFP expression was stable in mycelia and fluorescence was visible in transgenic fruiting bodies and GFP was detectable in planta. Use of these vectors has been successful in giving expression of the fluorescent proteins GFP and mRFP in A. mellea, providing an additional molecular tool for this pathogen.
DOI: 10.1038/srep29226
PubMed: 27384974
PubMed Central: PMC4935854
Affiliations:
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Foster</name><affiliation wicri:level="1"><nlm:affiliation>School of Biological Sciences, Life Sciences Building, University of Bristol, 24 Tyndall Avenue, Bristol, BS8 1TQ, United Kingdom.</nlm:affiliation><country xml:lang="fr">Royaume-Uni</country><wicri:regionArea>School of Biological Sciences, Life Sciences Building, University of Bristol, 24 Tyndall Avenue, Bristol, BS8 1TQ</wicri:regionArea><wicri:noRegion>BS8 1TQ</wicri:noRegion></affiliation></author></analytic><series><title level="j">Scientific reports</title><idno type="eISSN">2045-2322</idno><imprint><date when="2016" type="published">2016</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Agaricus (genetics)</term><term>Armillaria (genetics)</term><term>Green Fluorescent Proteins (genetics)</term><term>Inclusion Bodies (genetics)</term><term>Introns (genetics)</term><term>Mycelium (genetics)</term><term>Nuclear Proteins (genetics)</term><term>Phanerochaete (genetics)</term><term>Promoter Regions, Genetic (genetics)</term></keywords><keywords scheme="KwdFr" xml:lang="fr"><term>Agaricus (génétique)</term><term>Armillaria (génétique)</term><term>Corps d'inclusion (génétique)</term><term>Introns (génétique)</term><term>Mycelium (génétique)</term><term>Phanerochaete (génétique)</term><term>Protéines nucléaires (génétique)</term><term>Protéines à fluorescence verte (génétique)</term><term>Régions promotrices (génétique) (génétique)</term></keywords><keywords scheme="MESH" type="chemical" qualifier="genetics" xml:lang="en"><term>Green Fluorescent Proteins</term><term>Nuclear Proteins</term></keywords><keywords scheme="MESH" qualifier="genetics" xml:lang="en"><term>Agaricus</term><term>Armillaria</term><term>Inclusion Bodies</term><term>Introns</term><term>Mycelium</term><term>Phanerochaete</term><term>Promoter Regions, Genetic</term></keywords><keywords scheme="MESH" qualifier="génétique" xml:lang="fr"><term>Agaricus</term><term>Armillaria</term><term>Corps d'inclusion</term><term>Introns</term><term>Mycelium</term><term>Phanerochaete</term><term>Protéines nucléaires</term><term>Protéines à fluorescence verte</term><term>Régions promotrices (génétique)</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">Armillaria mellea is a significant pathogen that causes Armillaria root disease on numerous hosts in forests, gardens and agricultural environments worldwide. Using a yeast-adapted pCAMBIA0380 Agrobacterium vector, we have constructed a series of vectors for transformation of A. mellea, assembled using yeast-based recombination methods. These have been designed to allow easy exchange of promoters and inclusion of introns. The vectors were first tested by transformation into basidiomycete Clitopilus passeckerianus to ascertain vector functionality then used to transform A. mellea. We show that heterologous promoters from the basidiomycetes Agaricus bisporus and Phanerochaete chrysosporium that were used successfully to control the hygromycin resistance cassette were not able to support expression of mRFP or GFP in A. mellea. The endogenous A. mellea gpd promoter delivered efficient expression, and we show that inclusion of an intron was also required for transgene expression. GFP and mRFP expression was stable in mycelia and fluorescence was visible in transgenic fruiting bodies and GFP was detectable in planta. Use of these vectors has been successful in giving expression of the fluorescent proteins GFP and mRFP in A. mellea, providing an additional molecular tool for this pathogen.</div></front></TEI><pubmed><MedlineCitation Status="MEDLINE" Owner="NLM"><PMID Version="1">27384974</PMID><DateCompleted><Year>2018</Year><Month>05</Month><Day>18</Day></DateCompleted><DateRevised><Year>2019</Year><Month>01</Month><Day>11</Day></DateRevised><Article PubModel="Electronic"><Journal><ISSN IssnType="Electronic">2045-2322</ISSN><JournalIssue CitedMedium="Internet"><Volume>6</Volume><PubDate><Year>2016</Year><Month>07</Month><Day>07</Day></PubDate></JournalIssue><Title>Scientific reports</Title><ISOAbbreviation>Sci Rep</ISOAbbreviation></Journal><ArticleTitle>A native promoter and inclusion of an intron is necessary for efficient expression of GFP or mRFP in Armillaria mellea.</ArticleTitle><Pagination><MedlinePgn>29226</MedlinePgn></Pagination><ELocationID EIdType="doi" ValidYN="Y">10.1038/srep29226</ELocationID><Abstract><AbstractText>Armillaria mellea is a significant pathogen that causes Armillaria root disease on numerous hosts in forests, gardens and agricultural environments worldwide. Using a yeast-adapted pCAMBIA0380 Agrobacterium vector, we have constructed a series of vectors for transformation of A. mellea, assembled using yeast-based recombination methods. These have been designed to allow easy exchange of promoters and inclusion of introns. The vectors were first tested by transformation into basidiomycete Clitopilus passeckerianus to ascertain vector functionality then used to transform A. mellea. We show that heterologous promoters from the basidiomycetes Agaricus bisporus and Phanerochaete chrysosporium that were used successfully to control the hygromycin resistance cassette were not able to support expression of mRFP or GFP in A. mellea. The endogenous A. mellea gpd promoter delivered efficient expression, and we show that inclusion of an intron was also required for transgene expression. GFP and mRFP expression was stable in mycelia and fluorescence was visible in transgenic fruiting bodies and GFP was detectable in planta. Use of these vectors has been successful in giving expression of the fluorescent proteins GFP and mRFP in A. mellea, providing an additional molecular tool for this pathogen.</AbstractText></Abstract><AuthorList CompleteYN="Y"><Author ValidYN="Y"><LastName>Ford</LastName><ForeName>Kathryn L</ForeName><Initials>KL</Initials><AffiliationInfo><Affiliation>School of Biological Sciences, Life Sciences Building, University of Bristol, 24 Tyndall Avenue, Bristol, BS8 1TQ, United Kingdom.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Baumgartner</LastName><ForeName>Kendra</ForeName><Initials>K</Initials><AffiliationInfo><Affiliation>United States Department of Agriculture-Agricultural Research Service, 363 Hutchison Hall, University of California, One Shields Avenue, Davis, CA 95616, United States.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Henricot</LastName><ForeName>Béatrice</ForeName><Initials>B</Initials><AffiliationInfo><Affiliation>The Royal Horticultural Society, Wisley, Woking, Surrey, GU23 6QB, United Kingdom.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Bailey</LastName><ForeName>Andy M</ForeName><Initials>AM</Initials><AffiliationInfo><Affiliation>School of Biological Sciences, Life Sciences Building, University of Bristol, 24 Tyndall Avenue, Bristol, BS8 1TQ, United Kingdom.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Foster</LastName><ForeName>Gary D</ForeName><Initials>GD</Initials><AffiliationInfo><Affiliation>School of Biological Sciences, Life Sciences Building, University of Bristol, 24 Tyndall Avenue, Bristol, BS8 1TQ, United Kingdom.</Affiliation></AffiliationInfo></Author></AuthorList><Language>eng</Language><PublicationTypeList><PublicationType UI="D016428">Journal Article</PublicationType><PublicationType UI="D013485">Research Support, Non-U.S. Gov't</PublicationType><PublicationType UI="D013486">Research Support, U.S. Gov't, 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MajorTopicYN="Y">genetics</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D049452" MajorTopicYN="N">Green Fluorescent Proteins</DescriptorName><QualifierName UI="Q000235" MajorTopicYN="Y">genetics</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D002479" MajorTopicYN="N">Inclusion Bodies</DescriptorName><QualifierName UI="Q000235" MajorTopicYN="N">genetics</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D007438" MajorTopicYN="N">Introns</DescriptorName><QualifierName UI="Q000235" MajorTopicYN="Y">genetics</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D025282" MajorTopicYN="N">Mycelium</DescriptorName><QualifierName UI="Q000235" MajorTopicYN="N">genetics</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D009687" MajorTopicYN="N">Nuclear Proteins</DescriptorName><QualifierName UI="Q000235" MajorTopicYN="Y">genetics</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D020075" 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|wiki= Bois
|area= PhanerochaeteV1
|flux= Main
|étape= Exploration
|type= RBID
|clé= pubmed:27384974
|texte= A native promoter and inclusion of an intron is necessary for efficient expression of GFP or mRFP in Armillaria mellea.
}}
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