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  • Life and Medical Sciences  (2)
  • 1
    Digitale Medien
    Digitale Medien
    Chichester [u.a.] : Wiley-Blackwell
    Developmental Genetics 10 (1989), S. 210-219 
    ISSN: 0192-253X
    Schlagwort(e): Intron ; Alcohol dehydrogenase ; Enhancer ; Promoter ; Life and Medical Sciences ; Genetics
    Quelle: Wiley InterScience Backfile Collection 1832-2000
    Thema: Biologie
    Notizen: We have used in vitro mutagenesis and somatic transformation [Sofer and Martin, 1987a; Martin et al., 1986] to investigate the role of cis-acting sequences in the control of alcohol dehydrogenase gene expression in larvae of Drosophila melanogaster. Two sets of experiments were carried out. In the first, a series of aeletions were constructed in the region upstream of the proximal transcriptional start site. In the second, one or both introns were removed from within the structural gene. These constructs (on circular plasmids) were injected into Adh-null embryos and ADH activity was assayed in third instar larvae of the injected generation. The first set of experiments indicated that there are at least three distinct regulatory regions essential for larval activity located in the 5′ flanking region of the gene. One, in an area that includes the TATA box, was found to be necessary but not sufficient for larval ADH activity. Two others, further upstream, seemed to have enhancer-like properties because their absence could be compensated by a second copy of the Adh gene on the same plasmid molecule. The second set of experiments showed that neither the tis-sue distribution nor amount of ADH activity was affected by the removal of one or both introns from the Adh gene.
    Zusätzliches Material: 4 Ill.
    Materialart: Digitale Medien
    Bibliothek Standort Signatur Band/Heft/Jahr Verfügbarkeit
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  • 2
    ISSN: 0192-253X
    Schlagwort(e): Alcohol dehydrogenase ; silencer ; tissue-specific position effect ; Life and Medical Sciences ; Genetics
    Quelle: Wiley InterScience Backfile Collection 1832-2000
    Thema: Biologie
    Notizen: A transient expression assay has been used to investigate the cause of a tissuespecific position effect on Adh expression from a transgene insertion in Drosophila. A 15.4-kb genomic clone containing the 3.2-kb Adh insert along with flanking regions of genomic DNA is expressed in this assay in a tissue-specific pattern resembling the abnormal expression pattern of the position effect. The 3.2-kb Adh insert is expressed normally without the flanking sequences. A silencer element is located upstream of the Adh gene within a 2-kb fragment that acts in both orientations and at a distance of at least 6.5 kb from the larval Adh promoter to suppress ADH expression in a nontissue specific fashion. The DNA sequence of the 2-kb fragment indicates that it is a noncoding region. A 17-bp sequence is repeated within this region and may be associated with the silencer activity, since subclones from the 2-kb fragment, each containing one of the repeated regions, both retain full silencer activity. This silencer fails to suppress expression from an α1-tubulin promoter-LacZ fusion construct or an hsp70 promoter-Ach fusion construct. In addition to the silencer, another element is located downstream of the Adh gene that produces a higher level of anterior than posterior midgut expression. These results suggest that the 5′ silencer and the 3′ element act together to create the tissue specific pcsition effect characteristic of the GC-1 line. © 1994 Wiley-Liss, Inc.
    Zusätzliches Material: 5 Ill.
    Materialart: Digitale Medien
    Bibliothek Standort Signatur Band/Heft/Jahr Verfügbarkeit
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