构巢曲霉重组蛋白分泌的转录组分析及体内未折叠蛋白反应
Transcriptome analysis of recombinant protein secretion by Aspergillus nidulans and the unfolded-protein response in vivo.
作者信息
Sims Andrew H, Gent Manda E, Lanthaler Karin, Dunn-Coleman Nigel S, Oliver Stephen G, Robson Geoffrey D
机构信息
Faculty of Life Sciences, The University of Manchester, Oxford Road, Manchester M13 9PT, UK.
出版信息
Appl Environ Microbiol. 2005 May;71(5):2737-47. doi: 10.1128/AEM.71.5.2737-2747.2005.
Abstract
Filamentous fungi have a high capacity for producing large amounts of secreted proteins, a property that has been exploited for commercial production of recombinant proteins. However, the secretory pathway, which is key to the production of extracellular proteins, is rather poorly characterized in filamentous fungi compared to yeast. We report the effects of recombinant protein secretion on gene expression levels in Aspergillus nidulans by directly comparing a bovine chymosin-producing strain with its parental wild-type strain in continuous culture by using expressed sequence tag microarrays. This approach demonstrated more subtle and specific changes in gene expression than those observed when mimicking the effects of protein overproduction by using a secretion blocker. The impact of overexpressing a secreted recombinant protein more closely resembles the unfolded-protein response in vivo.
摘要
丝状真菌具有产生大量分泌蛋白的强大能力，这一特性已被用于重组蛋白的商业生产。然而，与酵母相比，丝状真菌中作为细胞外蛋白产生关键的分泌途径的特征还很不清楚。我们通过使用表达序列标签微阵列，在连续培养中直接比较产牛凝乳酶的菌株与其亲本野生型菌株，报告了重组蛋白分泌对构巢曲霉基因表达水平的影响。与使用分泌阻断剂模拟蛋白质过量产生的影响时所观察到的情况相比，这种方法证明了基因表达中更细微和特异的变化。过表达一种分泌型重组蛋白的影响更类似于体内的未折叠蛋白反应。
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