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里氏木霉营养型和孢子结合型疏水蛋白的差异表达——hfb2基因的克隆与表征

Differential expression of the vegetative and spore-bound hydrophobins of Trichoderma reesei--cloning and characterization of the hfb2 gene.

作者信息

Nakari-Setälä T, Aro N, Ilmén M, Muñoz G, Kalkkinen N, Penttilä M

机构信息

VTT Biotechnology and Food Research, Finland.

出版信息

Eur J Biochem. 1997 Sep 1;248(2):415-23. doi: 10.1111/j.1432-1033.1997.00415.x.

Abstract

The hfb2 gene encoding the hydrophobin HFBII of the filamentous fungus Trichoderma reesei was isolated by heterologous hybridization using the vegetative hydrophobin I, hfb1, gene of T. reesei as a probe. The hfb2 gene codes for a typical fungal secreted hydrophobin of 71 amino acids containing eight cysteine residues. The amino acid similarity towards HFBI is 69%. The HFBII protein was isolated from the fungal spores by extraction with trifluoroacetic acid/acetonitrile solution, and by bubbling from the lactose-based culture medium. Expression of the hfb1 and hfb2 genes is divergent. hfb1 expression was only observed in vegetative cultures on glucose-containing and sorbitol-containing media. It was not expressed on media containing complex plant polysaccharides, cellulose, xylan, cellobiose or lactose, whereas hfb2 was highly expressed in vegetative cultures on these media. Expression of hfb2 was also strongly induced by N and C starvation, by light and in conidiating cultures.

摘要

利用里氏木霉的营养疏水蛋白I(hfb1)基因作为探针,通过异源杂交分离出了编码丝状真菌里氏木霉疏水蛋白HFBII的hfb2基因。hfb2基因编码一种典型的真菌分泌型疏水蛋白,由71个氨基酸组成,含有8个半胱氨酸残基。与HFBI的氨基酸相似性为69%。通过用三氟乙酸/乙腈溶液提取以及从基于乳糖的培养基中鼓泡的方式,从真菌孢子中分离出了HFBII蛋白。hfb1和hfb2基因的表达是不同的。仅在含葡萄糖和含山梨醇的培养基上的营养培养物中观察到hfb1的表达。在含有复合植物多糖、纤维素、木聚糖、纤维二糖或乳糖的培养基上不表达,而hfb2在这些培养基上的营养培养物中高度表达。hfb2的表达也受到氮和碳饥饿、光照以及产孢培养物的强烈诱导。

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