Fowler T, Berka R M, Ward M
Genencor International, South San Francisco, CA 94080.
Curr Genet. 1990 Dec;18(6):537-45. doi: 10.1007/BF00327025.
The glucoamylase gene of Aspergillus niger, glaA, is expressed at high levels in the presence of starch. We have determined the nucleotide sequence of 1966 bp of the 5' flanking region of the glaA gene and have begun to identify sequences important for the control of glaA expression by deletion analysis. Constructs containing deletions extending into the glaA gene promoter were introduced into an A. niger host whose own glaA gene had been disrupted by a gene replacement event. Secreted levels of glucoamylase, expressed from each of the recombinant glaA genes, were measured by enzyme immunoassay. The effect of each deletion on the expression of glaA, when grown on differing carbon sources, was used to determine the limits of sequences upstream of glaA responsible for gene regulation. A region between -562 and -318 appears to direct high-level expression, whereas only 214 bp of 5' flanking sequence is required to initiate the start to transcription.
黑曲霉的糖化酶基因glaA在有淀粉存在的情况下高水平表达。我们已经测定了glaA基因5'侧翼区域1966 bp的核苷酸序列,并已开始通过缺失分析来鉴定对glaA表达控制重要的序列。将包含延伸到glaA基因启动子的缺失片段的构建体导入到一个黑曲霉宿主中,该宿主自身的glaA基因已通过基因置换事件被破坏。通过酶免疫测定法测量从每个重组glaA基因表达的糖化酶的分泌水平。当在不同碳源上生长时,每个缺失对glaA表达的影响被用于确定glaA上游负责基因调控的序列界限。-562至-318之间的区域似乎指导高水平表达,而启动转录起始仅需要214 bp的5'侧翼序列。