在体内和体外细菌蛋白表达系统之间的翻译启动一致性
Jiaojiao Li1,2, Peixian Li1,2, Qian Liu1,2
1School of Chemical Engineering and Technology, Tianjin University, Tianjin, China.
Frontiers in bioengineering and biotechnology
|June 12, 2023
概括
这项研究系统地分析了细菌5'未翻译区域 (5'-UTRs) 以获得一致的蛋白质翻译. 研究结果显示,更简单的缺乏核酸C的5'-UTR和复杂的结构在细胞和无细胞系统中提高了翻译效率.
科学领域:
- 合成生物学 合成生物学
- 分子生物学分子生物学
- 生物技术是生物技术.
背景情况:
- 5'-终端未翻译区域 (5'-UTR) 是调节细菌中翻译启动的关键.
- 在合成生物学中对遗传元素的标准化需要了解不同系统的5'-UTR功能一致性.
研究的目的:
- 系统地评估不同细菌菌株和体外系统中的5'-UTR介导蛋白转化的一致性.
- 确定影响翻译效率的5'-UTR的序列和结构特征.
主要方法:
- 描述了400多个表达卡塞特,其中包括各种调节GFP基因表达的5'-UTRs.
- 在两个大肠杆菌菌株 (JM109,BL21) 和一个无细胞体外系统中比较蛋白质翻译一致性.
- 对标准的统计热力学模型进行评估.
主要成果:
- 在两个体内细胞系统 (JM109和BL21) 之间观察到蛋白质翻译一致性的强烈相关性.
- 在体内和体外系统之间失去了蛋白质翻译一致性,两者都偏离了热力学模型.
- 缺少核酸C和减少5'-UTR中的二次结构显著提高了蛋白质翻译效率.
结论:
- 细菌的5'-UTR功能在体内表现出很高的一致性,但在体内无细胞系统中显著分离.
- 5'-UTRs的序列和结构特征,特别是核酸组成和二次结构,是翻译效率的关键决定因素.
- 确定了优化5'-UTR元件的设计原则,以在合成生物学应用中增强蛋白质合成.
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