Fluorescence microscopy: a powerful technique to detect low GUS activity in vascular tissues.
Identifieur interne : 004443 ( Main/Exploration ); précédent : 004442; suivant : 004444Fluorescence microscopy: a powerful technique to detect low GUS activity in vascular tissues.
Auteurs : Philippe Rech [France] ; Jacqueline Grima-Pettenati ; Alain JauneauSource :
- The Plant journal : for cell and molecular biology [ 0960-7412 ] ; 2003.
Descripteurs français
- KwdFr :
- Délétion de séquence (MeSH), Eucalyptus (cytologie), Eucalyptus (enzymologie), Eucalyptus (génétique), Glucuronidase (analyse), Glucuronidase (génétique), Glucuronidase (métabolisme), Histocytochimie (MeSH), Microscopie de fluorescence (méthodes), Mutagenèse (MeSH), Protéines de fusion recombinantes (analyse), Protéines de fusion recombinantes (métabolisme), Sensibilité et spécificité (MeSH).
- MESH :
- analyse : Glucuronidase, Protéines de fusion recombinantes.
- cytologie : Eucalyptus.
- enzymologie : Eucalyptus.
- génétique : Eucalyptus, Glucuronidase.
- métabolisme : Glucuronidase, Protéines de fusion recombinantes.
- méthodes : Microscopie de fluorescence.
- Délétion de séquence, Histocytochimie, Mutagenèse, Sensibilité et spécificité.
English descriptors
- KwdEn :
- Eucalyptus (cytology), Eucalyptus (enzymology), Eucalyptus (genetics), Glucuronidase (analysis), Glucuronidase (genetics), Glucuronidase (metabolism), Histocytochemistry (MeSH), Microscopy, Fluorescence (methods), Mutagenesis (MeSH), Recombinant Fusion Proteins (analysis), Recombinant Fusion Proteins (metabolism), Sensitivity and Specificity (MeSH), Sequence Deletion (MeSH).
- MESH :
- chemical , analysis : Glucuronidase, Recombinant Fusion Proteins.
- cytology : Eucalyptus.
- enzymology : Eucalyptus.
- genetics : Eucalyptus, Glucuronidase.
- chemical , metabolism : Glucuronidase, Recombinant Fusion Proteins.
- methods : Microscopy, Fluorescence.
- Histocytochemistry, Mutagenesis, Sensitivity and Specificity, Sequence Deletion.
Abstract
We have previously shown that the Eucalyptus gunnii EgCAD2 promoter was preferentially expressed in vascular tissues in different transgenic plants (poplar, tobacco, Arabidopsis and grapevine). In order to delineate the cis elements governing this vascular expression pattern, promoter deletion analysis was performed allowing us to identify the proximal region [-340/-124] as essential for vascular cambium/xylem-specific expression. In plants transformed with the smallest promoter region [-124/+117], the GUS activity was difficult to detect using conventional bright field microscopy. To overcome this problem, we used fluorescence microscopy, enabling us to show that the [-124/+117] region contained cis-elements driving activity in phloem fibres but not in secondary xylem. The technical improvement of the histochemical detection of GUS activity using fluorescence microscopy enables accurate investigation of low GUS activity in phenol-rich tissues.
DOI: 10.1046/j.1365-313x.2003.016017.x
PubMed: 12943553
Affiliations:
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(methods)</term><term>Mutagenesis (MeSH)</term><term>Recombinant Fusion Proteins (analysis)</term><term>Recombinant Fusion Proteins (metabolism)</term><term>Sensitivity and Specificity (MeSH)</term><term>Sequence Deletion (MeSH)</term></keywords><keywords scheme="KwdFr" xml:lang="fr"><term>Délétion de séquence (MeSH)</term><term>Eucalyptus (cytologie)</term><term>Eucalyptus (enzymologie)</term><term>Eucalyptus (génétique)</term><term>Glucuronidase (analyse)</term><term>Glucuronidase (génétique)</term><term>Glucuronidase (métabolisme)</term><term>Histocytochimie (MeSH)</term><term>Microscopie de fluorescence (méthodes)</term><term>Mutagenèse (MeSH)</term><term>Protéines de fusion recombinantes (analyse)</term><term>Protéines de fusion recombinantes (métabolisme)</term><term>Sensibilité et spécificité (MeSH)</term></keywords><keywords scheme="MESH" type="chemical" qualifier="analysis" xml:lang="en"><term>Glucuronidase</term><term>Recombinant Fusion Proteins</term></keywords><keywords 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In order to delineate the cis elements governing this vascular expression pattern, promoter deletion analysis was performed allowing us to identify the proximal region [-340/-124] as essential for vascular cambium/xylem-specific expression. In plants transformed with the smallest promoter region [-124/+117], the GUS activity was difficult to detect using conventional bright field microscopy. To overcome this problem, we used fluorescence microscopy, enabling us to show that the [-124/+117] region contained cis-elements driving activity in phloem fibres but not in secondary xylem. The technical improvement of the histochemical detection of GUS activity using fluorescence microscopy enables accurate investigation of low GUS activity in phenol-rich tissues.</div></front></TEI><pubmed><MedlineCitation Status="MEDLINE" Owner="NLM"><PMID Version="1">12943553</PMID><DateCompleted><Year>2003</Year><Month>11</Month><Day>07</Day></DateCompleted><DateRevised><Year>2019</Year><Month>09</Month><Day>06</Day></DateRevised><Article PubModel="Print"><Journal><ISSN IssnType="Print">0960-7412</ISSN><JournalIssue CitedMedium="Print"><Volume>33</Volume><Issue>1</Issue><PubDate><Year>2003</Year><Month>Jan</Month></PubDate></JournalIssue><Title>The Plant journal : for cell and molecular biology</Title><ISOAbbreviation>Plant J</ISOAbbreviation></Journal><ArticleTitle>Fluorescence microscopy: a powerful technique to detect low GUS activity in vascular tissues.</ArticleTitle><Pagination><MedlinePgn>205-9</MedlinePgn></Pagination><Abstract><AbstractText>We have previously shown that the Eucalyptus gunnii EgCAD2 promoter was preferentially expressed in vascular tissues in different transgenic plants (poplar, tobacco, Arabidopsis and grapevine). 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