在Trichoderma reesei中增强异质基因表达,通过促进多拷贝集成促进多拷贝集成
Hugues Mathis1, Delphine Naquin2, Antoine Margeot3
1IFP Energies Nouvelles, 1 et 4 Avenue de Bois-Préau, 92852, Rueil-Malmaison, France. hugues.mathis@ifpen.fr.
Applied microbiology and biotechnology
|September 23, 2024
概括
这项研究表明,Trichoderma reesei可以使用tef1促进体有效地产生异质蛋白质. 表达等离子体的多复制集成导致蛋白质的生产增加,优化这个真菌平台的工业应用.
科学领域:
- 生物技术是生物技术.
- 分子生物学分子生物学
- 微生物工程 微生物工程
背景情况:
- 特里科德尔马 (Trichoderma reesei) 是一种用于异种蛋白质生产的真菌平台.
- 之前的工作开发了使用tef1促进器和cbh1终止器的表达盒.
- 变形体显示了可变的光,表明不同的表达水平.
研究的目的:
- 调查Trichoderma reesei中异质基因的基因整合和表达.
- 了解等离子体拷贝数和蛋白质表达水平之间的关系.
- 为了优化Trichoderma reesei作为一个增强蛋白质生产的宿主.
主要方法:
- 开发了一个表达式磁带,其中有tef1促进器和cbh1终止器.
- 用含有egfp转基因的等离子体生成Trichoderma reesei转基因.
- 使用Illumina和牛津纳米孔平台进行全基因组测序.
- 分析等离子体融合部位和复制号码.
主要成果:
- 所有的转变体都显示出Tef1基因上游的等离子体集成,通过同源重组.
- 等离子体集成有利于基因组中的多拷贝集成.
- 较高的等离子体拷贝数与增加的EGFP光和蛋白质过度生产相关.
- tef1促进器和cbh1终止器系统显示了可变的eGFP表达水平.
结论:
- 基于tef1促进体的表达系统促进了Trichoderma reesei中的多拷贝集成和异构蛋白的过度生产.
- 这个系统是通过增加基因拷贝数来改善高价值蛋白质生产的宝贵工具.
- 优化基因整合策略可以提高Trichoderma reesei作为生产平台的效率.
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