McKenzie R W, Brennan M D
Department of Biochemistry and Molecular Biology, School of Medicine, University of Louisville, KY 40292, USA.
Insect Biochem Mol Biol. 1998 Nov;28(11):869-73. doi: 10.1016/s0965-1748(98)00072-1.
To characterize the cis-acting sequences required for expression of the alcohol dehydrogenase (Adh) gene of the Hawaiian picture-winged fruit fly Drosophila affinidisjuncta, germ-line transformation was used to introduce altered forms of this gene into Drosophila melanogaster. Genes were constructed lacking regions corresponding to known or putative regulatory elements within the D. melanogaster gene as well as to sequences previously shown to be required for expression of the D. affinidisjuncta gene in the larval fat body as assayed by transient transformation. Measurement of alcohol dehydrogenase (EC 1.1.1.1) activity levels produced by the transfected genes indicates that multiple elements in both the 5' and 3' regions of the gene contribute to expression. The dispersed elements appear to function redundantly to ensure high levels of expression. Moreover, in contrast to what has been reported for other Drosophila Adh genes, some of the regulatory elements influence expression in both larvae and adults. In total, these results reveal a regulatory system in which the transcribed region in imbedded in an extended genomic segment rich in cisacting regulatory information.
为了鉴定夏威夷图翅果蝇Drosophila affinidisjuncta乙醇脱氢酶(Adh)基因表达所需的顺式作用序列,采用种系转化法将该基因的变异形式导入黑腹果蝇。构建的基因缺失了黑腹果蝇基因内对应于已知或推定调控元件的区域,以及先前通过瞬时转化分析表明在幼虫脂肪体中D. affinidisjuncta基因表达所需的序列。对转染基因产生的乙醇脱氢酶(EC 1.1.1.1)活性水平的测量表明,该基因5'和3'区域中的多个元件对表达有贡献。这些分散的元件似乎以冗余方式发挥作用以确保高水平的表达。此外,与其他果蝇Adh基因的报道相反,一些调控元件影响幼虫和成虫两者的表达。总的来说,这些结果揭示了一种调控系统,其中转录区域嵌入在富含顺式作用调控信息的扩展基因组片段中。