Identification of a Bidirectional Promoter from <i>Trichoderma reesei</i> and Its Application in Dual Gene Expression
The cellulolytic filamentous fungus <i>Trichoderma reesei</i> has a strong capability in protein synthesis and secretion and is increasingly used as a fungal chassis for the production of heterologous proteins or secondary metabolites. However, bidirectional promoters that would signific...
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MDPI AG
2022-10-01
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| Series: | Journal of Fungi |
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| Online Access: | https://www.mdpi.com/2309-608X/8/10/1059 |
| _version_ | 1797472289742127104 |
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| author | Xiaoxiao Wu Fuzhe Li Renfei Yang Xiangfeng Meng Weixin Zhang Weifeng Liu |
| author_facet | Xiaoxiao Wu Fuzhe Li Renfei Yang Xiangfeng Meng Weixin Zhang Weifeng Liu |
| author_sort | Xiaoxiao Wu |
| collection | DOAJ |
| description | The cellulolytic filamentous fungus <i>Trichoderma reesei</i> has a strong capability in protein synthesis and secretion and is increasingly used as a fungal chassis for the production of heterologous proteins or secondary metabolites. However, bidirectional promoters that would significantly facilitate multiple genes’ expression have not been characterized in <i>T. reesei</i>. Herein, we show that a 767-bp intergenic region between two polyketide synthase encoding genes that were involved in the biosynthesis of the typical yellow pigment served as a bidirectional promoter in <i>T. reesei</i>. This region was shown to be able to drive the simultaneous expression of two fluorescence reporter genes when fused to each end. Quantitative RT-PCR analysis demonstrated that the driving strength of this bidirectional promoter from each direction reached about half of that of the commonly used promoter P<i>gpdA</i>. Moreover, the co-expression of two cellulase genes driven by this bidirectional promoter enabled <i>T. reesei</i> to produce cellulases on glucose and improved the total cellulase activities with cellulose Avicel as the carbon source. Our work identified the first bidirectional promoter in <i>T. reesei</i>, which would facilitate gene co-expression and find applications in synthetic biology using fungal systems. |
| first_indexed | 2024-03-09T19:59:45Z |
| format | Article |
| id | doaj.art-30de982a33df4b568b6ec0e18a27afe3 |
| institution | Directory Open Access Journal |
| issn | 2309-608X |
| language | English |
| last_indexed | 2024-03-09T19:59:45Z |
| publishDate | 2022-10-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Journal of Fungi |
| spelling | doaj.art-30de982a33df4b568b6ec0e18a27afe32023-11-24T00:47:20ZengMDPI AGJournal of Fungi2309-608X2022-10-01810105910.3390/jof8101059Identification of a Bidirectional Promoter from <i>Trichoderma reesei</i> and Its Application in Dual Gene ExpressionXiaoxiao Wu0Fuzhe Li1Renfei Yang2Xiangfeng Meng3Weixin Zhang4Weifeng Liu5State Key Laboratory of Microbial Technology, Shandong University, No.72 Binhai Road, Qingdao 266237, ChinaState Key Laboratory of Microbial Technology, Shandong University, No.72 Binhai Road, Qingdao 266237, ChinaState Key Laboratory of Microbial Technology, Shandong University, No.72 Binhai Road, Qingdao 266237, ChinaState Key Laboratory of Microbial Technology, Shandong University, No.72 Binhai Road, Qingdao 266237, ChinaState Key Laboratory of Microbial Technology, Shandong University, No.72 Binhai Road, Qingdao 266237, ChinaState Key Laboratory of Microbial Technology, Shandong University, No.72 Binhai Road, Qingdao 266237, ChinaThe cellulolytic filamentous fungus <i>Trichoderma reesei</i> has a strong capability in protein synthesis and secretion and is increasingly used as a fungal chassis for the production of heterologous proteins or secondary metabolites. However, bidirectional promoters that would significantly facilitate multiple genes’ expression have not been characterized in <i>T. reesei</i>. Herein, we show that a 767-bp intergenic region between two polyketide synthase encoding genes that were involved in the biosynthesis of the typical yellow pigment served as a bidirectional promoter in <i>T. reesei</i>. This region was shown to be able to drive the simultaneous expression of two fluorescence reporter genes when fused to each end. Quantitative RT-PCR analysis demonstrated that the driving strength of this bidirectional promoter from each direction reached about half of that of the commonly used promoter P<i>gpdA</i>. Moreover, the co-expression of two cellulase genes driven by this bidirectional promoter enabled <i>T. reesei</i> to produce cellulases on glucose and improved the total cellulase activities with cellulose Avicel as the carbon source. Our work identified the first bidirectional promoter in <i>T. reesei</i>, which would facilitate gene co-expression and find applications in synthetic biology using fungal systems.https://www.mdpi.com/2309-608X/8/10/1059bidirectional promoterfilamentous funguscellulase<i>sor</i> cluster |
| spellingShingle | Xiaoxiao Wu Fuzhe Li Renfei Yang Xiangfeng Meng Weixin Zhang Weifeng Liu Identification of a Bidirectional Promoter from <i>Trichoderma reesei</i> and Its Application in Dual Gene Expression Journal of Fungi bidirectional promoter filamentous fungus cellulase <i>sor</i> cluster |
| title | Identification of a Bidirectional Promoter from <i>Trichoderma reesei</i> and Its Application in Dual Gene Expression |
| title_full | Identification of a Bidirectional Promoter from <i>Trichoderma reesei</i> and Its Application in Dual Gene Expression |
| title_fullStr | Identification of a Bidirectional Promoter from <i>Trichoderma reesei</i> and Its Application in Dual Gene Expression |
| title_full_unstemmed | Identification of a Bidirectional Promoter from <i>Trichoderma reesei</i> and Its Application in Dual Gene Expression |
| title_short | Identification of a Bidirectional Promoter from <i>Trichoderma reesei</i> and Its Application in Dual Gene Expression |
| title_sort | identification of a bidirectional promoter from i trichoderma reesei i and its application in dual gene expression |
| topic | bidirectional promoter filamentous fungus cellulase <i>sor</i> cluster |
| url | https://www.mdpi.com/2309-608X/8/10/1059 |
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