Wu C Y, Brennan M D
Biochemistry Department, University of Louisville School of Medicine, Kentucky 40292.
Mol Gen Genet. 1993 Jul;240(1):58-64. doi: 10.1007/BF00276884.
In Drosophila melanogaster transformants, the alcohol dehydrogenase (Adh) genes from D. affinidisjuncta and D. grimshawi show similar levels of expression except in the adult midgut where the D. affinidisjuncta gene is expressed about 10- to 20-fold more strongly. To study the arrangement of cis-acting sequences responsible for this regulatory difference, homologous restriction sites were used to create a series of chimeric genes that switched fragments from the 5' and 3' flanking regions of these two genes. Chimeric genes were introduced into the germ-line of D. melanogaster, and Adh gene expression was analyzed by measuring RNA levels. Various gene fragments in the promoter region and elsewhere influence expression in the adult midgut and in whole larvae and adults. Comparison of these results with earlier studies involving chimeras between the D. affinidisjuncta and D. hawaiiensis genes indicates that expression in the adult midgut is influenced by multiple regulatory sequences and that distinct arrangements of regulatory sequences can result in similar levels of expression both in the adult midgut and in the whole organism.
在黑腹果蝇转化体中,来自亲缘远缘果蝇和格氏果蝇的乙醇脱氢酶(Adh)基因显示出相似的表达水平,但在成年中肠中除外,在成年中肠中,亲缘远缘果蝇基因的表达强度要高出约10至20倍。为了研究造成这种调控差异的顺式作用序列的排列方式,利用同源限制性位点构建了一系列嵌合基因,这些嵌合基因交换了这两个基因5'和3'侧翼区域的片段。将嵌合基因导入黑腹果蝇的种系,并通过测量RNA水平来分析Adh基因的表达。启动子区域和其他位置的各种基因片段会影响成年中肠以及整个幼虫和成虫中的表达。将这些结果与早期涉及亲缘远缘果蝇和夏威夷果蝇基因之间嵌合体的研究进行比较表明,成年中肠中的表达受多个调控序列影响,并且调控序列的不同排列方式可导致成年中肠和整个生物体中出现相似的表达水平。