Rapid isolation of promoter sequences by TAIL-PCR: the 5′-flanking regions of Pal and Pgi genes from yams ( Dioscorea )
Identifieur interne : 003702 ( Main/Exploration ); précédent : 003701; suivant : 003703Rapid isolation of promoter sequences by TAIL-PCR: the 5′-flanking regions of Pal and Pgi genes from yams ( Dioscorea )
Auteurs : R. Terauchi [États-Unis] ; G. Kahl [États-Unis]Source :
- Molecular and General Genetics MGG [ 0026-8925 ] ; 2000-04-01.
Descripteurs français
- KwdFr :
- Analyse de séquence d'ADN (), Données de séquences moléculaires, Exons, Glucose 6-phosphate isomerase (génétique), Introns, Liliaceae (génétique), Modèles génétiques, Phenylalanine ammonia-lyase (génétique), Réaction de polymérisation en chaîne (), Régions promotrices (génétique), Similitude de séquences d'acides nucléiques, Séquence nucléotidique.
- MESH :
- génétique : Glucose 6-phosphate isomerase, Liliaceae, Phenylalanine ammonia-lyase.
- Analyse de séquence d'ADN, Données de séquences moléculaires, Exons, Introns, Modèles génétiques, Réaction de polymérisation en chaîne, Régions promotrices (génétique), Similitude de séquences d'acides nucléiques, Séquence nucléotidique.
English descriptors
- KwdEn :
- Base Sequence, Dioscorea, Exons, Glucose-6-Phosphate Isomerase (genetics), Introns, Key words TAIL-PCR, Liliaceae (genetics), Models, Genetic, Molecular Sequence Data, PAL gene, PGI gene, Phenylalanine Ammonia-Lyase (genetics), Polymerase Chain Reaction (methods), Promoter, Promoter Regions, Genetic, Sequence Analysis, DNA (methods), Sequence Homology, Nucleic Acid.
- MESH :
- chemical , genetics : Glucose-6-Phosphate Isomerase, Phenylalanine Ammonia-Lyase.
- genetics : Liliaceae.
- methods : Polymerase Chain Reaction, Sequence Analysis, DNA.
- Base Sequence, Exons, Introns, Models, Genetic, Molecular Sequence Data, Promoter Regions, Genetic, Sequence Homology, Nucleic Acid.
Abstract
Abstract: Using a modified TAIL-PCR technique, the 5′-flanking regions of the phenylalanine ammonia lyase (Pal) genes of a yam species, Dioscorea bulbifera, and the phosphoglucose isomerase (Pgi) gene of D. tokoro were successfully isolated. Two novel modifications of the TAIL-PCR procedure introduced here, namely (1) the use of a battery of random 10-mers (RAPD primers) as short arbitrary primers, and (2) the use of a total of five nested, gene-specific primers, allow the rapid isolation of the 5′-flanking region of any gene from organisms with large genomes. Isolated 5′-flanking regions were fused to the gus gene, and tested for transient expression in tobacco BY2 cells. All the isolated 5′-flanking regions were shown to drive reporter gene expression. Three Pal promoters responded to salicylic acid, presumably as a result of the binding of a MYB transcriptional activator to the multiple MREs (Myb Recognition Elements) present in these regions.
Url:
DOI: 10.1007/s004380051201
Affiliations:
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Le document en format XML
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Two novel modifications of the TAIL-PCR procedure introduced here, namely (1) the use of a battery of random 10-mers (RAPD primers) as short arbitrary primers, and (2) the use of a total of five nested, gene-specific primers, allow the rapid isolation of the 5′-flanking region of any gene from organisms with large genomes. Isolated 5′-flanking regions were fused to the gus gene, and tested for transient expression in tobacco BY2 cells. All the isolated 5′-flanking regions were shown to drive reporter gene expression. Three Pal promoters responded to salicylic acid, presumably as a result of the binding of a MYB transcriptional activator to the multiple MREs (Myb Recognition Elements) present in these regions.</div></front></TEI><affiliations><list><country><li>États-Unis</li></country><region><li>Delaware</li></region></list><tree><country name="États-Unis"><region name="Delaware"><name sortKey="Terauchi, R" sort="Terauchi, R" uniqKey="Terauchi R" first="R." last="Terauchi">R. Terauchi</name></region><name sortKey="Kahl, G" sort="Kahl, G" uniqKey="Kahl G" first="G." last="Kahl">G. Kahl</name></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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