| Improved heterologous gene expression in Trichoderma reesei by cellobiohydrolase I gene （cbh1） promoter optimization |
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| 作者姓名： | Liu T Wang T Li X Liu X |
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| 作者单位： | State Key Laboratory of Microbial Technology, Shandong University, Jinan 250100, China |
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| 基金项目： | This work was supported by the grants from the National Natural Science Foundation of China （No. 30470052）, the National Basic Research Program （973） of China （No. 2003CB716006 and 2004CB719702） and the Natural Science Research Foundation for the Doctoral Program of the Higher Education Ministry of China （No. 20040422042） |
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| 摘 要： | To improve heterologous gene expression in Trichoderma reesei, a set of optimal artificial cellobiohydrolase I gene （cbhl） promoters was obtained. The region from -677 to -724 with three potential glucose repressor binding sites was deleted. Then the region from -620 to -820 of the modified cbhl promoter, including the CCAAT box and the Ace2 binding site, was repeatedly inserted into the modified cbhl promoter, obtaining promoters with copy numbers 2, 4, and 6. The results showed that the glucose repression effects were abolished and the expression level of the glucuronidase （gus） reporter gene regulated by these multi-copy promoters was markedly enhanced as the copy number increased simultaneously. The data showed the great promise of using the promoter artificial modification strategy to increase heterologous gene expression in filamentous fungi and provided a set of optional high-expression vectors for gene function investigation and strain modification.
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| 关 键 词： | 助催化剂 目标缺失 基因 遗传学 |
| 收稿时间： | 2007-10-25 |
| 修稿时间： | 2007-12-13 |
Improved heterologous gene expression in Trichoderma reesei by cellobiohydrolase I gene (cbh1) promoter optimization |
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| Liu T,Wang T,Li X,Liu X.Improved heterologous gene expression in Trichoderma reesei by cellobiohydrolase I gene (cbh1) promoter optimization[J].Acta Biochimica et Biophysica Sinica,2008,40(2):158-165. |
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| Authors: | Liu Ti Wang Tianhong Li Xian Liu Xuan |
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| Institution: | State Key Laboratory of Microbial Technology, Shandong University, Jinan 250100, China |
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| Abstract: | To improve heterologous gene expression in Trichoderma reesei , a set of optimal artificial cellobiohydrolase I gene ( cbh1 ) promoters was obtained. The region from -677 to -724 with three potential glucose repressor binding sites was deleted. Then the region from -620 to -820 of the modified cbh1 promoter, including the CCAAT box and the Ace2 binding site, was repeatedly inserted into the modified cbh1 promoter, obtaining promoters with copy numbers 2, 4, and 6. The results showed that the glucose repression effects were abolished and the expression level of the glucuronidase ( gus ) reporter gene regulated by these multi-copy promoters was markedly enhanced as the copy number increased simultaneously. The data showed the great promise of using the promoter artificial modification strategy to increase heterologous gene expression in filamentous fungi and provided a set of optional high-expression vectors for gene function investigation and strain modification. |
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| Keywords: | Trichoderma reesei cbhl promoter carboncatabolite derepression targeted deletion multiple-copystrategy |
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