Transient Transactivation Studies in Nicotiana benthamiana Leaves.
Identifieur interne : 000151 ( Main/Exploration ); précédent : 000150; suivant : 000152Transient Transactivation Studies in Nicotiana benthamiana Leaves.
Auteurs : Pilar Lasierra [Espagne] ; Salomé Prat [Espagne]Source :
- Methods in molecular biology (Clifton, N.J.) [ 1940-6029 ] ; 2018.
Descripteurs français
- KwdFr :
- Activation de la transcription (MeSH), Agrobacterium (MeSH), Extinction de l'expression des gènes (MeSH), Feuilles de plante (génétique), Feuilles de plante (métabolisme), Luciferases (génétique), Luciferases (métabolisme), Protéines végétales (génétique), Protéines végétales (métabolisme), Régions promotrices (génétique) (MeSH), Régulation de l'expression des gènes végétaux (MeSH), Tabac (génétique), Tabac (métabolisme), Transcription génétique (MeSH), Végétaux génétiquement modifiés (génétique), Végétaux génétiquement modifiés (métabolisme).
- MESH :
- génétique : Feuilles de plante, Luciferases, Protéines végétales, Tabac, Végétaux génétiquement modifiés.
- métabolisme : Feuilles de plante, Luciferases, Protéines végétales, Tabac, Végétaux génétiquement modifiés.
- Activation de la transcription, Agrobacterium, Extinction de l'expression des gènes, Régions promotrices (génétique), Régulation de l'expression des gènes végétaux, Transcription génétique.
English descriptors
- KwdEn :
- Agrobacterium (MeSH), Gene Expression Regulation, Plant (MeSH), Gene Silencing (MeSH), Luciferases (genetics), Luciferases (metabolism), Plant Leaves (genetics), Plant Leaves (metabolism), Plant Proteins (genetics), Plant Proteins (metabolism), Plants, Genetically Modified (genetics), Plants, Genetically Modified (metabolism), Promoter Regions, Genetic (MeSH), Tobacco (genetics), Tobacco (metabolism), Transcription, Genetic (MeSH), Transcriptional Activation (MeSH).
- MESH :
- chemical , genetics : Luciferases, Plant Proteins.
- chemical , metabolism : Luciferases, Plant Proteins.
- genetics : Plant Leaves, Plants, Genetically Modified, Tobacco.
- metabolism : Plant Leaves, Plants, Genetically Modified, Tobacco.
- Agrobacterium, Gene Expression Regulation, Plant, Gene Silencing, Promoter Regions, Genetic, Transcription, Genetic, Transcriptional Activation.
Abstract
Transitory gene expression systems in Nicotiana benthamiana leaves, in combination with the use of gene silencing suppressors as the p19 or HC-pro proteins that allow for elevated levels of gene expression, have proven to be a highly versatile tool to analyze transcriptional function of DNA binding factors in the activated or repressed expression of their gene targets. This experimental setup uses Agrobacterium-mediated infection to deliver the various DNA constructs into the cell, and offers the advantage with respect to mesophyll protoplast transfection procedures that it entails a much easier protocol, in addition to preserving the intact leaf tissue, thus being more amenable to the study of wound and stress signaling pathways or to the functional analyses of regulators that respond to Ca+2 signatures. Furthermore, by using reporter constructs based on the LUCIFERASE (LUC) gene, which does not require a destructive determination assay, this expression system can be used to test for changes in gene activity over time or in response to various treatments, thus providing a comprehensive understanding of the signaling pathways that modulate activity of the expressed regulators and therefore their in vivo function in the control of the analyzed promoter.
DOI: 10.1007/978-1-4939-7871-7_22
PubMed: 29855968
Affiliations:
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Genética Molecular de Plantas, Centro Nacional de Biotecnología-CSIC, Madrid, Spain. sprat@cnb.csic.es.</nlm:affiliation><country xml:lang="fr">Espagne</country><wicri:regionArea>Dpto. 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Genética Molecular de Plantas, Centro Nacional de Biotecnología-CSIC, Madrid, Spain.</nlm:affiliation><country xml:lang="fr">Espagne</country><wicri:regionArea>Dpto. Genética Molecular de Plantas, Centro Nacional de Biotecnología-CSIC, Madrid</wicri:regionArea><placeName><settlement type="city">Madrid</settlement><region nuts="2" type="region">Communauté de Madrid</region></placeName></affiliation></author><author><name sortKey="Prat, Salome" sort="Prat, Salome" uniqKey="Prat S" first="Salomé" last="Prat">Salomé Prat</name><affiliation wicri:level="3"><nlm:affiliation>Dpto. Genética Molecular de Plantas, Centro Nacional de Biotecnología-CSIC, Madrid, Spain. sprat@cnb.csic.es.</nlm:affiliation><country xml:lang="fr">Espagne</country><wicri:regionArea>Dpto. Genética Molecular de Plantas, Centro Nacional de Biotecnología-CSIC, Madrid</wicri:regionArea><placeName><settlement type="city">Madrid</settlement><region nuts="2" type="region">Communauté de Madrid</region></placeName></affiliation></author></analytic><series><title level="j">Methods in molecular biology (Clifton, N.J.)</title><idno type="eISSN">1940-6029</idno><imprint><date when="2018" type="published">2018</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Agrobacterium (MeSH)</term><term>Gene Expression Regulation, Plant (MeSH)</term><term>Gene Silencing (MeSH)</term><term>Luciferases (genetics)</term><term>Luciferases (metabolism)</term><term>Plant Leaves (genetics)</term><term>Plant Leaves (metabolism)</term><term>Plant Proteins (genetics)</term><term>Plant Proteins (metabolism)</term><term>Plants, Genetically Modified (genetics)</term><term>Plants, Genetically Modified (metabolism)</term><term>Promoter Regions, Genetic (MeSH)</term><term>Tobacco (genetics)</term><term>Tobacco (metabolism)</term><term>Transcription, Genetic (MeSH)</term><term>Transcriptional Activation (MeSH)</term></keywords><keywords scheme="KwdFr" xml:lang="fr"><term>Activation de la transcription (MeSH)</term><term>Agrobacterium (MeSH)</term><term>Extinction de l'expression des gènes (MeSH)</term><term>Feuilles de plante (génétique)</term><term>Feuilles de plante (métabolisme)</term><term>Luciferases (génétique)</term><term>Luciferases (métabolisme)</term><term>Protéines végétales (génétique)</term><term>Protéines végétales (métabolisme)</term><term>Régions promotrices (génétique) (MeSH)</term><term>Régulation de l'expression des gènes végétaux (MeSH)</term><term>Tabac (génétique)</term><term>Tabac (métabolisme)</term><term>Transcription génétique (MeSH)</term><term>Végétaux génétiquement modifiés (génétique)</term><term>Végétaux génétiquement modifiés (métabolisme)</term></keywords><keywords scheme="MESH" type="chemical" qualifier="genetics" xml:lang="en"><term>Luciferases</term><term>Plant Proteins</term></keywords><keywords scheme="MESH" type="chemical" qualifier="metabolism" xml:lang="en"><term>Luciferases</term><term>Plant Proteins</term></keywords><keywords scheme="MESH" qualifier="genetics" xml:lang="en"><term>Plant Leaves</term><term>Plants, Genetically Modified</term><term>Tobacco</term></keywords><keywords scheme="MESH" qualifier="génétique" xml:lang="fr"><term>Feuilles de plante</term><term>Luciferases</term><term>Protéines végétales</term><term>Tabac</term><term>Végétaux génétiquement modifiés</term></keywords><keywords scheme="MESH" qualifier="metabolism" xml:lang="en"><term>Plant Leaves</term><term>Plants, Genetically Modified</term><term>Tobacco</term></keywords><keywords scheme="MESH" qualifier="métabolisme" xml:lang="fr"><term>Feuilles de plante</term><term>Luciferases</term><term>Protéines végétales</term><term>Tabac</term><term>Végétaux génétiquement modifiés</term></keywords><keywords scheme="MESH" xml:lang="en"><term>Agrobacterium</term><term>Gene Expression Regulation, Plant</term><term>Gene Silencing</term><term>Promoter Regions, Genetic</term><term>Transcription, Genetic</term><term>Transcriptional Activation</term></keywords><keywords scheme="MESH" xml:lang="fr"><term>Activation de la transcription</term><term>Agrobacterium</term><term>Extinction de l'expression des gènes</term><term>Régions promotrices (génétique)</term><term>Régulation de l'expression des gènes végétaux</term><term>Transcription génétique</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">Transitory gene expression systems in Nicotiana benthamiana leaves, in combination with the use of gene silencing suppressors as the p19 or HC-pro proteins that allow for elevated levels of gene expression, have proven to be a highly versatile tool to analyze transcriptional function of DNA binding factors in the activated or repressed expression of their gene targets. This experimental setup uses Agrobacterium-mediated infection to deliver the various DNA constructs into the cell, and offers the advantage with respect to mesophyll protoplast transfection procedures that it entails a much easier protocol, in addition to preserving the intact leaf tissue, thus being more amenable to the study of wound and stress signaling pathways or to the functional analyses of regulators that respond to Ca<sup>+2</sup> signatures. Furthermore, by using reporter constructs based on the LUCIFERASE (LUC) gene, which does not require a destructive determination assay, this expression system can be used to test for changes in gene activity over time or in response to various treatments, thus providing a comprehensive understanding of the signaling pathways that modulate activity of the expressed regulators and therefore their in vivo function in the control of the analyzed promoter.</div></front></TEI><pubmed><MedlineCitation Status="MEDLINE" Owner="NLM"><PMID Version="1">29855968</PMID><DateCompleted><Year>2019</Year><Month>02</Month><Day>25</Day></DateCompleted><DateRevised><Year>2019</Year><Month>02</Month><Day>25</Day></DateRevised><Article PubModel="Print"><Journal><ISSN IssnType="Electronic">1940-6029</ISSN><JournalIssue CitedMedium="Internet"><Volume>1794</Volume><PubDate><Year>2018</Year></PubDate></JournalIssue><Title>Methods in molecular biology (Clifton, N.J.)</Title><ISOAbbreviation>Methods Mol Biol</ISOAbbreviation></Journal><ArticleTitle>Transient Transactivation Studies in Nicotiana benthamiana Leaves.</ArticleTitle><Pagination><MedlinePgn>311-322</MedlinePgn></Pagination><ELocationID EIdType="doi" ValidYN="Y">10.1007/978-1-4939-7871-7_22</ELocationID><Abstract><AbstractText>Transitory gene expression systems in Nicotiana benthamiana leaves, in combination with the use of gene silencing suppressors as the p19 or HC-pro proteins that allow for elevated levels of gene expression, have proven to be a highly versatile tool to analyze transcriptional function of DNA binding factors in the activated or repressed expression of their gene targets. This experimental setup uses Agrobacterium-mediated infection to deliver the various DNA constructs into the cell, and offers the advantage with respect to mesophyll protoplast transfection procedures that it entails a much easier protocol, in addition to preserving the intact leaf tissue, thus being more amenable to the study of wound and stress signaling pathways or to the functional analyses of regulators that respond to Ca<sup>+2</sup> signatures. Furthermore, by using reporter constructs based on the LUCIFERASE (LUC) gene, which does not require a destructive determination assay, this expression system can be used to test for changes in gene activity over time or in response to various treatments, thus providing a comprehensive understanding of the signaling pathways that modulate activity of the expressed regulators and therefore their in vivo function in the control of the analyzed promoter.</AbstractText></Abstract><AuthorList CompleteYN="Y"><Author ValidYN="Y"><LastName>Lasierra</LastName><ForeName>Pilar</ForeName><Initials>P</Initials><AffiliationInfo><Affiliation>Dpto. Genética Molecular de Plantas, Centro Nacional de Biotecnología-CSIC, Madrid, Spain.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Prat</LastName><ForeName>Salomé</ForeName><Initials>S</Initials><AffiliationInfo><Affiliation>Dpto. Genética Molecular de Plantas, Centro Nacional de Biotecnología-CSIC, Madrid, Spain. sprat@cnb.csic.es.</Affiliation></AffiliationInfo></Author></AuthorList><Language>eng</Language><PublicationTypeList><PublicationType UI="D016428">Journal Article</PublicationType></PublicationTypeList></Article><MedlineJournalInfo><Country>United States</Country><MedlineTA>Methods Mol Biol</MedlineTA><NlmUniqueID>9214969</NlmUniqueID><ISSNLinking>1064-3745</ISSNLinking></MedlineJournalInfo><ChemicalList><Chemical><RegistryNumber>0</RegistryNumber><NameOfSubstance UI="D010940">Plant Proteins</NameOfSubstance></Chemical><Chemical><RegistryNumber>EC 1.13.12.-</RegistryNumber><NameOfSubstance UI="D008156">Luciferases</NameOfSubstance></Chemical></ChemicalList><CitationSubset>IM</CitationSubset><MeshHeadingList><MeshHeading><DescriptorName UI="D060054" MajorTopicYN="N">Agrobacterium</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D018506" MajorTopicYN="Y">Gene Expression Regulation, Plant</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D020868" MajorTopicYN="N">Gene Silencing</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D008156" MajorTopicYN="N">Luciferases</DescriptorName><QualifierName UI="Q000235" MajorTopicYN="N">genetics</QualifierName><QualifierName UI="Q000378" MajorTopicYN="N">metabolism</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D018515" MajorTopicYN="N">Plant Leaves</DescriptorName><QualifierName UI="Q000235" MajorTopicYN="N">genetics</QualifierName><QualifierName UI="Q000378" MajorTopicYN="Y">metabolism</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D010940" MajorTopicYN="N">Plant Proteins</DescriptorName><QualifierName UI="Q000235" MajorTopicYN="N">genetics</QualifierName><QualifierName UI="Q000378" MajorTopicYN="Y">metabolism</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D030821" MajorTopicYN="N">Plants, Genetically Modified</DescriptorName><QualifierName UI="Q000235" MajorTopicYN="N">genetics</QualifierName><QualifierName UI="Q000378" MajorTopicYN="Y">metabolism</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D011401" MajorTopicYN="N">Promoter Regions, Genetic</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D014026" MajorTopicYN="N">Tobacco</DescriptorName><QualifierName UI="Q000235" MajorTopicYN="N">genetics</QualifierName><QualifierName UI="Q000378" MajorTopicYN="Y">metabolism</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D014158" MajorTopicYN="N">Transcription, Genetic</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D015533" MajorTopicYN="Y">Transcriptional Activation</DescriptorName></MeshHeading></MeshHeadingList><KeywordList Owner="NOTNLM"><Keyword MajorTopicYN="Y">Agroinfiltration</Keyword><Keyword MajorTopicYN="Y">Conserved regulatory element</Keyword><Keyword MajorTopicYN="Y">Luciferase activity</Keyword><Keyword MajorTopicYN="Y">Promoter region</Keyword><Keyword MajorTopicYN="Y">Silencing suppressor</Keyword><Keyword MajorTopicYN="Y">Transcription factor</Keyword></KeywordList></MedlineCitation><PubmedData><History><PubMedPubDate PubStatus="entrez"><Year>2018</Year><Month>6</Month><Day>2</Day><Hour>6</Hour><Minute>0</Minute></PubMedPubDate><PubMedPubDate PubStatus="pubmed"><Year>2018</Year><Month>6</Month><Day>2</Day><Hour>6</Hour><Minute>0</Minute></PubMedPubDate><PubMedPubDate PubStatus="medline"><Year>2019</Year><Month>2</Month><Day>26</Day><Hour>6</Hour><Minute>0</Minute></PubMedPubDate></History><PublicationStatus>ppublish</PublicationStatus><ArticleIdList><ArticleId IdType="pubmed">29855968</ArticleId><ArticleId IdType="doi">10.1007/978-1-4939-7871-7_22</ArticleId></ArticleIdList></PubmedData></pubmed><affiliations><list><country><li>Espagne</li></country><region><li>Communauté de Madrid</li></region><settlement><li>Madrid</li></settlement></list><tree><country name="Espagne"><region name="Communauté de Madrid"><name sortKey="Lasierra, Pilar" sort="Lasierra, Pilar" uniqKey="Lasierra P" first="Pilar" last="Lasierra">Pilar Lasierra</name></region><name sortKey="Prat, Salome" sort="Prat, Salome" uniqKey="Prat S" first="Salomé" last="Prat">Salomé Prat</name></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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