Identification of a Genomic Region Containing a Novel Promoter Resistant to Glucose Repression and Over-Expression of beta-Glucosidase Gene in Hypocrea orientalis EU7-22

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	Identification of a Genomic Region Containing a Novel Promoter Resistant to Glucose Repression and Over-Expression of beta-Glucosidase Gene in Hypocrea orientalis EU7-22
	Long, Chuannan ; Cheng, Yijin ; Gan, Lihui ; Liu, Jian ; Long, Minnan ; Long MN(龙敏南)
刊名	http://dx.doi.org/10.3390/ijms14048479
	2013
关键词	AGROBACTERIUM-MEDIATED TRANSFORMATION FUNGUS TRICHODERMA-REESEI ASPERGILLUS-NIDULANS BINDING-SITES TUMEFACIENS WALKING CLONING TOOL
英文摘要	National Basic Research Program of China (973 Program) [2010CB732201]; Fundamental Research Funds for the Central Universities of China [201112G026]; National Natural Science Foundation of China [31170067]; A high concentration of glucose in the medium could greatly inhibit the expression of cellulase in filamentous fungi. The aspartic protease from fungus Hypocrea orientalis EU7-22 could efficiently express under both induction condition and glucose repression condition. Based on the sequence of structure gene of aspartic protease, the upstream sequence harboring the putative promoter proA for driving the expression of aspartic protease was obtained by genome walking. The upstream sequence contained the typical promoter motifs "TATA" and "CAAT". The beta-glucosidase gene (Bgl1) from H. orientalis was cloned and recombined with promoter proA and terminator trpC. The expression cassette was ligated to the binary vector to form pUR5750-Bgl1, and then transferred into the host strain EU7-22 via Agrobacterium tumefaciens mediated transformation (ATMT), using hygromycin B resistance gene as the screening marker. Four transformants Bgl-1, Bgl-2, Bgl-3 and Bgl-4 were screened. Compared with the host strain EU7-22, the enzyme activities of filter paper (FPA) and beta-glucosidase (BG) of transformant Bgl-2 increased by 10.6% and 19.1% under induction condition, respectively. The FPA and BG activities were enhanced by 22.2% and 700% under 2% glucose repression condition, respectively, compared with the host strain. The results showed that the putative promoter proA has successfully driven the over-expression of Bgl1 gene in H. orientalis under glucose repression condition.
语种	英语
出版者	MDPI AG
内容类型	期刊论文
源URL	[http://dspace.xmu.edu.cn/handle/2288/90735]
专题	生命科学－已发表论文
推荐引用方式 GB/T 7714	Long, Chuannan,Cheng, Yijin,Gan, Lihui,et al. Identification of a Genomic Region Containing a Novel Promoter Resistant to Glucose Repression and Over-Expression of beta-Glucosidase Gene in Hypocrea orientalis EU7-22[J]. http://dx.doi.org/10.3390/ijms14048479,2013.
APA	Long, Chuannan,Cheng, Yijin,Gan, Lihui,Liu, Jian,Long, Minnan,&龙敏南.(2013).Identification of a Genomic Region Containing a Novel Promoter Resistant to Glucose Repression and Over-Expression of beta-Glucosidase Gene in Hypocrea orientalis EU7-22.http://dx.doi.org/10.3390/ijms14048479.
MLA	Long, Chuannan,et al."Identification of a Genomic Region Containing a Novel Promoter Resistant to Glucose Repression and Over-Expression of beta-Glucosidase Gene in Hypocrea orientalis EU7-22".http://dx.doi.org/10.3390/ijms14048479 (2013).