文献类型：期刊文献

英文题名：A light-switchable bidirectional expression system in filamentous fungus Trichoderma reesei

作者：Zhang, Guoxiu[1];Liu, Pei[1];Wei, Wei[1];Wang, Xuedong[1];Wei, Dongzhi[1];Wang, Wei[1]

机构：[1]East China Univ Sci & Technol, New World Inst Biotechnol, State Key Lab Bioreactor Engn, Shanghai 200237, Peoples R China

年份：2016

卷号：240

起止页码：85

外文期刊名：JOURNAL OF BIOTECHNOLOGY

收录：;EI(收录号：20164603007961);WOS:【SCI-EXPANDED(收录号:WOS:000389116700010)】；

基金：This research was supported by the National Natural Science Foundation of China (No. C010302-31500066), the National Basic Research Program of China (973, Program No. 2012CB721103), and theNational High Technology Research and Development Program of China (863, Program No. 2013AA102109).

语种：英文

外文关键词：Trichoderma reesei; Light-switchable system; Heterologous protein; Simultaneous expression; Cellulase-free background

摘要：The filamentous fungi Trichoderma reesei is widely used in the production of cellulolytic enzymes and recombinant proteins. However, only moderate success has been achieved in expressing heterologous proteins in T. reesei. Light-dependent control of DNA transcription, and protein expression have been demonstrated in bacteria, fungi, and mammalian cells. In this study, light inducible transactivators, a "light-on" bidirectional promoter and a "light-off" promoter were constructed successfully in T. reesei for the first time. Our light inducible transactivators can homodimerize and bind to the upstream region of artificial promoters to activate or repress genes transcription. Additionally, we upgraded the light-inducible system to on-off system that can simultaneously control the expression of multiple heterologous proteins in T. reesei. Moreover, a native cellulase-free background for the expression of heterologous proteins was achieved by knocking out the genes involved in transcriptional regulation and encoding of cellulases: xyr1, cbh1, and cbh2. Our light-switchable system showed a very little background protein expression and robust activation in the blue light with significantly improved heterologous protein expression. We demonstrate that our light-switchable system has a potential application as an on/off "switch" that can simultaneously regulate the expression of multiple genes in T. reesei under native cellulase-free background. (C) 2016 Elsevier B.V. All rights reserved.