Deletion scanning of the regulatory sequences of the Fbp1 gene of Drosophila melanogaster using P transposase-induced deficiencies.

Abstract:

: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.

journal_name

Genetics

journal_title

Genetics

authors

Lapie P,Nasr F,Lepesant JA,Deutsch J

subject

Has Abstract

pub_date

1993-11-01 00:00:00

pages

801-16

issue

3

eissn

0016-6731

issn

1943-2631

journal_volume

135

pub_type

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