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在基因组尺度上对5'UTRs的核酸分布分析指导它们在酵母中的重新设计和表达调节

Chaoying Yao¹, Yu Yin², Qingyang Li¹

¹State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, 130 Meilong Road, Shanghai, 200237, China.

Metabolic engineering

|December 29, 2024

概括

研究人员通过分析5'未翻译区域 (5'UTRs) 在非传统酵母中发现了新的翻译控制工具. 修改特定序列,特别是KZ3基因,可显著增强工业应用中的基因表达.

科学领域:

分子生物学分子生物学
合成生物学合成生物学
生物技术是生物技术.

背景情况:

非传统酵母对生物工业产品有价值.
对于控制这些酵母中基因表达的工具有限.
翻译性调节机制经常被忽视.

研究的目的:

识别和描述酵母转化调节的新型工具.
为了增强非传统酵母宿主中的基因表达控制.

主要方法:

在四种酵母物种中对5'未翻译区域 (5'UTR) 的基因组规模分析.
通过基替代和核酸三重组偏好的分析 (5'UTRs的再生 (KZ3动机).
使用不同的促进剂 (P_AOX1,P_GAP) 构建和测试KZ3变体.
多基因组分析以确定基因表达增强的水平.

主要成果:

在 Komagataella phaffii 5'UTRs 的 -100 到 -1 区域中,发现了"G"频率的下降.
在 -50 到 -1 区域中用"A"或"T"取代"G"显著增强了基因表达.
发现KZ3基因 (-3至-1区域) 主要影响基因表达.
在128种构造的KZ3变体中,58种增加了基因表达,差异高达15倍.
增强发生在翻译,而不是转录层面.

结论:

基于5'UTR的新型工具,特别是KZ3图案,可以微调酵母体的转化调节.
使用这些工具实现了工业蛋白质和平台化合物的改善生产.
这项研究为增强酵母和其他真菌的基因表达提供了宝贵的参考资料和工具.

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