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ooc1,一种仅在嗜热栖热放线菌(无性型:里氏木霉)在纤维素上生长期间表达的独特基因。

ooc1, a unique gene expressed only during growth of Hypocrea jecorina (anamorph: Trichoderma reesei) on cellulose.

作者信息

Schmoll Monika, Kubicek Christian P

机构信息

Division of Gene Technology and Applied Biochemistry, Institute for Chemical Engineering, Vienna University of Technology, Austria.

出版信息

Curr Genet. 2005 Aug;48(2):126-33. doi: 10.1007/s00294-005-0585-1. Epub 2005 Sep 14.

Abstract

To grow on cellulose as a carbon source, Hypocrea jecorina (Trichoderma reesei) expresses and secretes a number of cellulases. This mechanism of induction by an insoluble carbon source has been controversially explained, but is most frequently attributed to the formation of the beta-1,2-diglucoside sophorose, a powerful soluble inducer of cellulases, by means of transglycosylation by constitutive or conidia-bound beta-glycoside hydrolases. Some recent results, however, have put the role of sophorose as the mediator of cellulose induction in question. Here we used the rapid subtraction hybridization approach to clone genes expressed by H. jecorina in the presence of cellulose but not upon incubation with sophorose. From a total of 96 expressed sequence tag (EST) fragments, 37 putative positives--representing ten different genes--were selected and analysed. All of them were present in the genome sequence of H. jecorina. Three of them encode proteins known from H. jecorina, five encode enzymes involved in secondary metabolism and one gene encodes an as yet unknown member of glycoside hydrolase family 30. Two EST fragments had no orthologues in other fungi. One of them made up for 25 of the 37 EST fragments analysed. The corresponding gene (only expressed on cellulose, ooc1) encodes a small secreted 10.5-kDa protein. The ooc1 transcript is only detectable during growth on cellulose in darkness, but not on cellulose in light or in the presence of other cellulase inducers (sophorose, lactose), nor is it formed during growth on glucose or glycerol. Its expression is strongly reduced, but not completely abolished in the cellulase non-inducible mutant QM 9978. The results of this study provide evidence that induction of gene expression by cellulose does not necessarily correlate with that by sophorose.

摘要

为了以纤维素作为碳源生长,嗜热栖热菌(里氏木霉)会表达并分泌多种纤维素酶。这种由不溶性碳源诱导的机制一直存在争议,但最常被认为是通过组成型或分生孢子结合的β-糖苷水解酶进行转糖基化作用,形成β-1,2-二葡萄糖苷槐糖,一种强大的纤维素酶可溶性诱导剂。然而,最近的一些结果对槐糖作为纤维素诱导介质的作用提出了质疑。在这里,我们使用快速消减杂交方法克隆嗜热栖热菌在存在纤维素但与槐糖孵育时不表达的基因。从总共96个表达序列标签(EST)片段中,选择并分析了37个推定的阳性片段——代表10个不同的基因。它们都存在于嗜热栖热菌的基因组序列中。其中三个编码嗜热栖热菌中已知的蛋白质,五个编码参与次级代谢的酶,一个基因编码糖苷水解酶家族30中一个未知成员。两个EST片段在其他真菌中没有直系同源物。其中一个占分析的37个EST片段中的25个。相应的基因(仅在纤维素上表达,ooc1)编码一种分泌的10.​​5 kDa小蛋白。ooc1转录本仅在黑暗中在纤维素上生长时可检测到,而在光照下在纤维素上或在存在其他纤维素酶诱导剂(槐糖、乳糖)时不可检测到,在葡萄糖或甘油上生长时也不形成。其表达在纤维素酶不可诱导的突变体QM 9978中强烈降低,但并未完全消除。这项研究的结果提供了证据,表明纤维素诱导的基因表达不一定与槐糖诱导的基因表达相关。

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