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一种基于泡盛曲霉1,4-β-内切木聚糖酶A基因启动子区域的表达系统。

An expression system based on the promoter region of the Aspergillus awamori 1,4-beta-endoxylanase A gene.

作者信息

Gouka R J, Hessing J G, Punt P J, Stam H, Musters W, Van den Hondel C A

机构信息

TNO Nutrition and Food Research Institute, Department of Molecular Genetics and Gene Technology, Rijswijk, The Netherlands.

出版信息

Appl Microbiol Biotechnol. 1996 Aug;46(1):28-35. doi: 10.1007/s002530050779.

Abstract

A new, highly inducible fungal promoter derived from the Aspergillus awamori 1,4-beta-endoxylanase A (exlA) gene is described. Induction analysis, carried out with the wild-type strain in shake flasks, showed that exlA expression in regulated at the transcriptional level. Using a beta-glucuronidase (uidA) reporter strategy, D-xylose was shown to be an efficient inducer of the exlA promoter, whereas sucrose or maltodextrin were not. Upon D-xylose induction, the exlA promoter was threefold more efficient than the frequently used A. niger glucoamylase (glaA) promoter under maltodextrin induction. Detailed induction analyses demonstrated that induction was dependent on the presence of D-xylose in the medium. Carbon-source-limited chemostat cultures with the uidA reporter strain showed that D-xylose was also a very good inducer in a fermenter, even in the presence of sucrose.

摘要

本文描述了一种源自泡盛曲霉1,4-β-内切木聚糖酶A(exlA)基因的新型、高诱导性真菌启动子。在摇瓶中对野生型菌株进行的诱导分析表明,exlA的表达在转录水平受到调控。使用β-葡萄糖醛酸酶(uidA)报告策略表明,D-木糖是exlA启动子的有效诱导剂,而蔗糖或麦芽糊精则不是。在D-木糖诱导下,exlA启动子的效率比在麦芽糊精诱导下常用的黑曲霉糖化酶(glaA)启动子高三倍。详细的诱导分析表明,诱导依赖于培养基中D-木糖的存在。使用uidA报告菌株进行的碳源限制恒化器培养表明,即使在存在蔗糖的情况下,D-木糖在发酵罐中也是一种非常好的诱导剂。

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