Deletion scanning of the regulatory sequences of the Fbp1 gene of Drosophila melanogaster using P transposase-induced deficiencies
| Autor: | J.-A. Lepesant, Fahd Nasr, J. Deutsch, P. Lapie |
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| Rok vydání: | 1993 |
| Předmět: |
Male
Transposable element Sequence analysis Molecular Sequence Data Negative regulatory element Transposases Genes Insect Investigations Biology P element Genes Reporter Genes Regulator Genetics Animals Drosophila Proteins Deletion mapping Crosses Genetic Transposase DNA Primers Sequence Deletion Reporter gene Base Sequence Alcohol Dehydrogenase Membrane Proteins Nuclear Proteins DNA Cadherins Nucleotidyltransferases DNA-Binding Proteins Drosophila melanogaster Mutagenesis Regulatory sequence Insect Hormones Female Transcription Factors |
| Zdroj: | Genetics. 135:801-816 |
| ISSN: | 1943-2631 |
| DOI: | 10.1093/genetics/135.3.801 |
| Popis: | A procedure permitting deletion scanning of potential cis-regulatory sequences within a transgene whose genomic position remains fixed was applied to the study of the upstream sequences of the ecdysteroid-inducible Fat-body-protein-1 (Fbp1) gene. Deficiencies were induced in a Fbp1:Adh fusion transgene by means of a secondary P transposase mutagenesis. Phenotypic and molecular screens were used to select mutant transposons that retained their original genomic location and carried a deletion affecting the Fbp1 sequences but not the Adh reporter gene. Molecular mapping of the deletion breakpoints was achieved by sequence analysis and expression of the reporter gene was quantified by measurement of ADH activity. This procedure was efficient in detecting cis-acting elements, even those with moderate effects on levels of gene expression. For example, we have succeeded in identifying a negative regulatory element. Deletion of this element leads to a 50% increase in the reporter ADH activity. This element binds the transcription factor AEF-1. In addition, we have detected a strong, positively acting element contained within a 32-bp region located immediately upstream of an ecdysone-response element. |
| Databáze: | OpenAIRE |
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