Isolation of a Polyubiquitin Promoter and Its Expression in Transgenic Potato Plants
| Autor: | T. Oosumi, W. R. Belknap, Joan E. Garbarino |
|---|---|
| Rok vydání: | 1995 |
| Předmět: |
clone (Java method)
Untranslated region Physiology Recombinant Fusion Proteins Molecular Sequence Data Gene Expression Plant Science Biology Genes Plant Polymerase Chain Reaction Biopolymers Complementary DNA Gene expression Genetics Genomic library Codon Polyubiquitin Promoter Regions Genetic Ubiquitins Gene DNA Primers Glucuronidase Solanum tuberosum Genomic Library Base Sequence fungi Intron food and beverages Agrobacterium tumefaciens Plants Genetically Modified biology.organism_classification Molecular biology Introns Kinetics Research Article |
| Zdroj: | Plant Physiology. 109:1371-1378 |
| ISSN: | 1532-2548 0032-0889 |
| Popis: | A polyubiquitin clone (ubi7) was isolated from a potato (Solanum tuberosum) genomic library using a copy-specific probe from a stress-induced ubiquitin cDNA. The genomic clone contained a 569-bp intron immediately 5[prime] to the initiation codon for the first ubiquitin-coding unit. Two chimeric [beta]-glucuronidase (GUS) fusion transgenes were introduced into potato. The first contained GUS fused to a 1156-bp promoter fragment containing only 5[prime] flanking and 5[prime] untranslated sequences from ubi7. The second transgene contained GUS translationally fused to the carboxy terminus of the first ubiquitin-coding unit and thus included the intron present in the 5[prime] untranslated region of the polyubiquitin gene. Both ubi7-GUS transgenes were activated by wounding in tuber tissue and in leaves by application of exogenous methyl jasmonate. They were also expressed constitutively in the potato tuber peel (outer 1–2 mm). Both transgenes were actively expressed in mature leaves. Exceptionally high levels of expression were observed in senescent leaves. Transgenic clones containing the ubi7 intron and the first ubiquitin-coding unit showed GUS expression levels at least 10 times higher than clones containing GUS fused to the intronless promoter. |
| Databáze: | OpenAIRE |
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