Analysis of the lupin Nodulin-45 promoter: conserved regulatory sequences are important for promoter activity
| Autor: | Kevin J. F. Farnden, Richard C. Macknight, Paul H. S. Reynolds |
|---|---|
| Rok vydání: | 1995 |
| Předmět: |
DNA
Plant Agrobacterium Recombinant Fusion Proteins Response element Molecular Sequence Data Plant Science Genes Plant Plant Roots Gene Expression Regulation Plant Genetics Promoter Regions Genetic Gene Conserved Sequence Plant Proteins Cell Nucleus Reporter gene Plants Medicinal biology Base Sequence Sequence Homology Amino Acid Membrane Proteins Promoter Fabaceae General Medicine biology.organism_classification Plants Genetically Modified Molecular biology DNA-Binding Proteins Transformation (genetics) Regulatory sequence Trans-acting Agronomy and Crop Science |
| Zdroj: | Plant molecular biology. 27(3) |
| ISSN: | 0167-4412 |
| Popis: | The promoter from the Lupinus angustifolius late nodulin gene, Nodulin-45, has been analysed to identify cis-elements and trans-acting factors. Various regions of the Nodulin-45 promoter, fused to the luciferase reporter gene, were introduced into Lotus roots using an Agrobacterium rhizogenes, transformation procedure. The transgenic roots were then nodulated. The promoter region A (-172 to +13, relative to the transcription start site) was capable of directing low-level expression of the reporter gene and in a nodule-enhanced manner when compared to roots. The addition of region C (-676 to -345) resulted in a significant increase in the expression within the nodule, whilst a low level of root expression was maintained. The C region, which confers this high-level nodule expression, contains the nodule consensus motifs AAAGAT and CTCTT. When region C was ligated to a minimal promoter element from the unrelated asparaginase gene rather than the Nodulin-45 A region, nodule-enhanced expression was still apparent, but at a much lower level. Mutation of the AAAGAT element in this construct resulted in a further significant decrease of expression. Gel retardation assays revealed that a factor from lupin nodule nuclear extracts interacted with two sequences of the C region. The binding of the factor to both of these regions could be removed by the addition of an oligonucleotide containing the AT-rich binding site for the soybean factor NAT2. This suggests that the lupin factor identified here is a NAT2 homologue. No factor binding was observed to the AAAGAT or CTCTT elements present in the C region. |
| Databáze: | OpenAIRE |
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