通过定点诱变上调米曲霉木聚糖酶基因的启动子活性

Upregulation of promoter activity of the Aspergillus oryzae xylanase gene by site-directed mutagenesis.

作者信息

Marui Junichiro, Kato Masashi, Kobayashi Tetsuo, Tsukagoshi Norihiro

机构信息

Department of Biological Mechanisms and Functions, Graduate School of Bioagricultural Sciences, Nagoya University, Nagoya 464-8601, Japan.

出版信息

Biotechnol Lett. 2003 Feb;25(4):371-4. doi: 10.1023/a:1022393130612.

DOI:10.1023/a:1022393130612

PMID:12882555

Abstract

In Aspergillus oryzae, inductive expression of the xynF1 gene is mediated by a transcriptional activator, AoXlnR. Promoter activity of the xynF1 gene, monitored by beta-galactosidase activity, was successfully upregulated by mutating two non-canonical AoXlnR binding sequences to what is thought to be the strongest sequence. Transformants carrying three canonical binding sequences in the promoter region produced enzyme 2.8 times more active (33,000 units mg(-1) protein) than that of transformants carrying the authentic promoter (12,000 units mg(-1) protein).

摘要

在米曲霉中，木聚糖酶基因xynF1的诱导表达由转录激活因子AoXlnR介导。通过β-半乳糖苷酶活性监测xynF1基因的启动子活性，将两个非典型的AoXlnR结合序列突变为据认为最强的序列后，启动子活性成功上调。在启动子区域携带三个典型结合序列的转化体产生的酶活性（33,000单位mg(-1)蛋白质）是携带原始启动子的转化体（12,000单位mg(-1)蛋白质）的2.8倍。