Hawley S A, Doane W W, Norman R A
Department of Zoology, Arizona State University, Tempe 85287-1501.
Biochem Genet. 1992 Jun;30(5-6):257-77.
The Amylase locus in Drosophila melanogaster contains duplicate, divergently transcribed structural genes for alpha-amylase, AmyA and AmyB. A sensitive and reliable transient expression assay was developed for testing amylase activities produced by exogenous Amy genes in somatically transformed larvae of an amylase-null strain of flies. Alleles tested, AmyA and AmyB, came from recombinant clone lambda Dm65, which contains genomic DNA from a Canton-S strain. The transient assay was used in a deletion analysis aimed at locating cis-regulatory sequences within the 5' region of AmyB. Results suggest that upstream regulatory sequences for correct spatial expression of AmyA and AmyB in third-instar larvae are located within 446 and 430 bp of their respective starts for transcription. A sequence required for high levels of AmyB expression was located within its 5' upstream region between the base pairs at -332 and -219. AmyA does not appear to have a comparable regulatory element in its 5'-flanking sequence. Barely detectable expression of AmyB was observed when it was flanked by only 92 bp of upstream sequence. A model is proposed for incomplete coordinate control of the duplicate Amy genes.
黑腹果蝇的淀粉酶基因座包含α-淀粉酶的重复、反向转录的结构基因AmyA和AmyB。开发了一种灵敏可靠的瞬时表达测定法,用于检测淀粉酶缺失型果蝇品系的体细胞转化幼虫中外源Amy基因产生的淀粉酶活性。所测试的等位基因AmyA和AmyB来自重组克隆λDm65,其包含来自Canton-S品系的基因组DNA。瞬时测定法用于缺失分析,旨在定位AmyB 5'区域内的顺式调控序列。结果表明,AmyA和AmyB在三龄幼虫中正确空间表达的上游调控序列位于其各自转录起始点的446和430 bp范围内。AmyB高水平表达所需的序列位于其5'上游区域中-332至-219碱基对之间。AmyA在其5'侧翼序列中似乎没有类似的调控元件。当AmyB仅侧翼有92 bp的上游序列时,观察到其表达几乎检测不到。提出了一个关于重复的Amy基因不完全协同调控的模型。