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鉴定和表征来自 Geosmithia spp. 的新型 II 类疏水蛋白 GEO1

Identification and characterization of GEO1, a new class II hydrophobin from Geosmithia spp.

机构信息

Dipartimento di Biologia Evoluzionistica "Leo Pardi", University of Florence, via Romana 17-19, 50125 Florence, Italy.

出版信息

Can J Microbiol. 2012 Aug;58(8):965-72. doi: 10.1139/w2012-069. Epub 2012 Jul 18.

Abstract

In the present paper we describe a new noncatalytic protein belonging to the hydrophobin family, designated GEO1, purified from the culture filtrate of Geosmithia pallida (Ascomycota: Hypocreales), and the corresponding gene sequence. In the fungal genome, GEO1 was encoded by a single-copy gene with a 450 bp open reading frame interrupted by 2 small introns whose primary translation product was 109 amino acids long and included a 23 amino acids signal peptide. The mature protein had a molecular mass of 8111.75 Da and a theoretical pI of 4.33. The deduced amino acid sequence showed similarity to class II hydrophobins and contained 8 conserved cysteine residues, present in all hydrophobins isolated so far. Biochemical properties, such as foam-forming ability and trapezoid-like shape of a GEO1 drop, also resembled the typical features of the class II hydrophobins. Expression of the geo1 gene was assessed after 2, 4, 7, 9, and 11 days of culture and showed that the geo1 transcript appeared after 7 days and increased up to 11 days.

摘要

在本文中,我们描述了一种新的非催化性蛋白质,属于疏水蛋白家族,命名为 GEO1,从 Geosmithia pallida(子囊菌门:Hypocreales)的培养滤液中纯化而来,并介绍了相应的基因序列。在真菌基因组中,GEO1 由一个单拷贝基因编码,该基因具有 450 bp 的开放阅读框,被 2 个小内含子打断,其主要翻译产物为 109 个氨基酸,包含一个 23 个氨基酸的信号肽。成熟蛋白的分子量为 8111.75 Da,理论等电点为 4.33。推导的氨基酸序列与 II 类疏水蛋白相似,含有 8 个保守的半胱氨酸残基,这些残基存在于迄今为止分离的所有疏水蛋白中。GEO1 液滴的成泡能力和梯形形状等生化特性也类似于 II 类疏水蛋白的典型特征。geo1 基因的表达在培养 2、4、7、9 和 11 天后进行评估,结果表明 geo1 转录本在第 7 天出现,并在第 11 天增加。

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