PoplarV1, Main, Exploration, bibRecord, 004477

Characterization of a PttRPS18 promoter active in the vascular cambium region of hybrid aspen.

Identifieur interne : 004477 ( Main/Exploration ); précédent : 004476; suivant : 004478

Characterization of a PttRPS18 promoter active in the vascular cambium region of hybrid aspen.

Auteurs : Ann-Marie Johansson [Suède] ; Chongying Wang ; Anneli Stenberg ; Magnus Hertzberg ; C H Anthony Little ; Olof Olsson

Source :

Plant molecular biology [ 0167-4412 ] ; 2003.

RBID : pubmed:12856939

Descripteurs français

English descriptors

KwdEn :
- Amino Acid Sequence (MeSH), Base Sequence (MeSH), Cloning, Molecular (MeSH), DNA, Bacterial (genetics), DNA, Complementary (chemistry), DNA, Complementary (genetics), DNA, Complementary (isolation & purification), DNA, Plant (chemistry), DNA, Plant (genetics), Gene Expression Regulation, Plant (MeSH), Glucuronidase (genetics), Glucuronidase (metabolism), Hybridization, Genetic (MeSH), Luciferases (genetics), Luciferases (metabolism), Molecular Sequence Data (MeSH), Plant Stems (genetics), Plant Stems (metabolism), Plant Structures (genetics), Plant Structures (metabolism), Plants, Genetically Modified (MeSH), Populus (genetics), Populus (metabolism), Promoter Regions, Genetic (genetics), RNA, Plant (genetics), RNA, Plant (metabolism), Recombinant Fusion Proteins (genetics), Recombinant Fusion Proteins (metabolism), Ribosomal Proteins (genetics), Sequence Analysis, DNA (MeSH), Transformation, Genetic (MeSH).
MESH :
- chemical , chemistry : DNA, Complementary, DNA, Plant.
- chemical , genetics : DNA, Bacterial, DNA, Complementary, DNA, Plant, Glucuronidase, Luciferases, RNA, Plant, Recombinant Fusion Proteins, Ribosomal Proteins.
- chemical , isolation & purification : DNA, Complementary.
- chemical , metabolism : Glucuronidase, Luciferases, RNA, Plant, Recombinant Fusion Proteins.
- genetics : Plant Stems, Plant Structures, Populus, Promoter Regions, Genetic.
- metabolism : Plant Stems, Plant Structures, Populus.
- Amino Acid Sequence, Base Sequence, Cloning, Molecular, Gene Expression Regulation, Plant, Hybridization, Genetic, Molecular Sequence Data, Plants, Genetically Modified, Sequence Analysis, DNA, Transformation, Genetic.

Abstract

By screening 273 hybrid aspen plants transformed with a luciferase-based promoter trap T-DNA vector, one plant was found in which the reporter gene (luxF2) was activated only in cells of the cambial region, i.e., vascular cambium, phloem and differentiating xylem. Southern blot analysis revealed that this plant denoted #24 had a single T-DNA insert. The chromosomal regions flanking the T-DNA were cloned by plasmid rescue. A 757 bp DNA fragment, originating from the rescued plasmid and covering the genomic region immediately upstream from the right-border sequence of the T-DNA, was used as a probe to isolate the corresponding chromosomal region from a wild-type hybrid aspen genomic library. A hybrid aspen small ribosomal protein gene, PttRPS18, was then isolated. By screening a wt cambial region-specific cDNA library, two cDNA clones encoding a putative 152 amino acid PttRPS18 protein were isolated. Comparison of the DNA sequence immediately flanking the T-DNA insert in #24 with the corresponding wild-type sequence showed that only a minor deletion occurred during the T-DNA integration. Northern analysis revealed that the PttRPS18 gene was expressed mainly in the cambial region. By RT-PCR and DNA sequencing analysis, the exact structures of the PttRPS18 and luxF2 transcripts were determined. Finally, the hybrid aspen PttRPS18 promoter was fused to the uidA reporter gene and transformed into hybrid aspen plants. Histochemical analysis of GUS activity showed that the PttRPS18 promoter was expressed in the cambial region in the same manner as the luciferase reporter gene in the initial screening.

DOI: 10.1023/a:1023919331037
PubMed: 12856939

Affiliations:

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Le document en format XML

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<term>Base Sequence (MeSH)</term>
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<term>DNA, Complementary (chemistry)</term>
<term>DNA, Complementary (genetics)</term>
<term>DNA, Complementary (isolation & purification)</term>
<term>DNA, Plant (chemistry)</term>
<term>DNA, Plant (genetics)</term>
<term>Gene Expression Regulation, Plant (MeSH)</term>
<term>Glucuronidase (genetics)</term>
<term>Glucuronidase (metabolism)</term>
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<term>Luciferases (genetics)</term>
<term>Luciferases (metabolism)</term>
<term>Molecular Sequence Data (MeSH)</term>
<term>Plant Stems (genetics)</term>
<term>Plant Stems (metabolism)</term>
<term>Plant Structures (genetics)</term>
<term>Plant Structures (metabolism)</term>
<term>Plants, Genetically Modified (MeSH)</term>
<term>Populus (genetics)</term>
<term>Populus (metabolism)</term>
<term>Promoter Regions, Genetic (genetics)</term>
<term>RNA, Plant (genetics)</term>
<term>RNA, Plant (metabolism)</term>
<term>Recombinant Fusion Proteins (genetics)</term>
<term>Recombinant Fusion Proteins (metabolism)</term>
<term>Ribosomal Proteins (genetics)</term>
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<term>ADN complémentaire (génétique)</term>
<term>ADN complémentaire (isolement et purification)</term>
<term>ADN des plantes (composition chimique)</term>
<term>ADN des plantes (génétique)</term>
<term>ARN des plantes (génétique)</term>
<term>ARN des plantes (métabolisme)</term>
<term>Analyse de séquence d'ADN (MeSH)</term>
<term>Clonage moléculaire (MeSH)</term>
<term>Données de séquences moléculaires (MeSH)</term>
<term>Glucuronidase (génétique)</term>
<term>Glucuronidase (métabolisme)</term>
<term>Hybridation génétique (MeSH)</term>
<term>Luciferases (génétique)</term>
<term>Luciferases (métabolisme)</term>
<term>Populus (génétique)</term>
<term>Populus (métabolisme)</term>
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<term>Protéines de fusion recombinantes (métabolisme)</term>
<term>Protéines ribosomiques (génétique)</term>
<term>Régions promotrices (génétique) (génétique)</term>
<term>Régulation de l'expression des gènes végétaux (MeSH)</term>
<term>Structures de plante (génétique)</term>
<term>Structures de plante (métabolisme)</term>
<term>Séquence d'acides aminés (MeSH)</term>
<term>Séquence nucléotidique (MeSH)</term>
<term>Tiges de plante (génétique)</term>
<term>Tiges de plante (métabolisme)</term>
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<term>DNA, Plant</term>
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<term>Populus</term>
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<term>ADN complémentaire</term>
<term>ADN des plantes</term>
<term>ARN des plantes</term>
<term>Glucuronidase</term>
<term>Luciferases</term>
<term>Populus</term>
<term>Protéines de fusion recombinantes</term>
<term>Protéines ribosomiques</term>
<term>Régions promotrices (génétique)</term>
<term>Structures de plante</term>
<term>Tiges de plante</term>
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<term>Plant Structures</term>
<term>Populus</term>
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<term>Luciferases</term>
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<term>Protéines de fusion recombinantes</term>
<term>Structures de plante</term>
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<term>Gene Expression Regulation, Plant</term>
<term>Hybridization, Genetic</term>
<term>Molecular Sequence Data</term>
<term>Plants, Genetically Modified</term>
<term>Sequence Analysis, DNA</term>
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<term>Données de séquences moléculaires</term>
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<front><div type="abstract" xml:lang="en">By screening 273 hybrid aspen plants transformed with a luciferase-based promoter trap T-DNA vector, one plant was found in which the reporter gene (luxF2) was activated only in cells of the cambial region, i.e., vascular cambium, phloem and differentiating xylem. Southern blot analysis revealed that this plant denoted #24 had a single T-DNA insert. The chromosomal regions flanking the T-DNA were cloned by plasmid rescue. A 757 bp DNA fragment, originating from the rescued plasmid and covering the genomic region immediately upstream from the right-border sequence of the T-DNA, was used as a probe to isolate the corresponding chromosomal region from a wild-type hybrid aspen genomic library. A hybrid aspen small ribosomal protein gene, PttRPS18, was then isolated. By screening a wt cambial region-specific cDNA library, two cDNA clones encoding a putative 152 amino acid PttRPS18 protein were isolated. Comparison of the DNA sequence immediately flanking the T-DNA insert in #24 with the corresponding wild-type sequence showed that only a minor deletion occurred during the T-DNA integration. Northern analysis revealed that the PttRPS18 gene was expressed mainly in the cambial region. By RT-PCR and DNA sequencing analysis, the exact structures of the PttRPS18 and luxF2 transcripts were determined. Finally, the hybrid aspen PttRPS18 promoter was fused to the uidA reporter gene and transformed into hybrid aspen plants. Histochemical analysis of GUS activity showed that the PttRPS18 promoter was expressed in the cambial region in the same manner as the luciferase reporter gene in the initial screening.</div>
</front>
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<DateCompleted><Year>2003</Year>
<Month>08</Month>
<Day>05</Day>
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<Month>08</Month>
<Day>23</Day>
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<Month>May</Month>
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<Title>Plant molecular biology</Title>
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<Abstract><AbstractText>By screening 273 hybrid aspen plants transformed with a luciferase-based promoter trap T-DNA vector, one plant was found in which the reporter gene (luxF2) was activated only in cells of the cambial region, i.e., vascular cambium, phloem and differentiating xylem. Southern blot analysis revealed that this plant denoted #24 had a single T-DNA insert. The chromosomal regions flanking the T-DNA were cloned by plasmid rescue. A 757 bp DNA fragment, originating from the rescued plasmid and covering the genomic region immediately upstream from the right-border sequence of the T-DNA, was used as a probe to isolate the corresponding chromosomal region from a wild-type hybrid aspen genomic library. A hybrid aspen small ribosomal protein gene, PttRPS18, was then isolated. By screening a wt cambial region-specific cDNA library, two cDNA clones encoding a putative 152 amino acid PttRPS18 protein were isolated. Comparison of the DNA sequence immediately flanking the T-DNA insert in #24 with the corresponding wild-type sequence showed that only a minor deletion occurred during the T-DNA integration. Northern analysis revealed that the PttRPS18 gene was expressed mainly in the cambial region. By RT-PCR and DNA sequencing analysis, the exact structures of the PttRPS18 and luxF2 transcripts were determined. Finally, the hybrid aspen PttRPS18 promoter was fused to the uidA reporter gene and transformed into hybrid aspen plants. Histochemical analysis of GUS activity showed that the PttRPS18 promoter was expressed in the cambial region in the same manner as the luciferase reporter gene in the initial screening.</AbstractText>
</Abstract>
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Characterization of a PttRPS18 promoter active in the vascular cambium region of hybrid aspen.

Characterization of a PttRPS18 promoter active in the vascular cambium region of hybrid aspen.

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