人与子混合翻译系统,探索修改后的核糖体的功能
Eriko Matsuura-Suzuki1, Hirotaka Toh1, Shintaro Iwasaki1,2
1RNA Systems Biochemistry Laboratory, RIKEN Cluster for Pioneering Research, Wako, Saitama 351-0198, Japan.
Bio-protocol
|July 17, 2023
概括
一种新的混合体体内翻译系统使用在耗尽的子网细胞溶解酸中净化的人类核糖体. 这种系统使得研究转化机制,特别是核糖体功能和过程性,克服了以前的分析障碍.
科学领域:
- 生物化学 生物化学
- 分子生物学分子生物学
- 翻译条例 翻译条例
背景情况:
- 在体外翻译系统对于研究翻译调节至关重要.
- 哺乳动物细胞提取物用于翻译是很难准备的;子网状细胞溶解物 (RRL) 是一个常见的替代品.
- 在RRL中分析核糖体功能存在分析挑战.
研究的目的:
- 开发一种混合翻译系统,通过补充纯化人类核糖体到核糖体贫乏的RRL.
- 为这种混合系统提供一个逐步的协议.
- 为了能够研究由缺乏翻译后修饰的核糖体驱动的翻译,并评估翻译延长过程性.
主要方法:
- 从子网状细胞溶解酸 (RRL) 来消耗内源性核糖体.
- 将净化的人类核糖体补充到耗尽的RRL中.
- 使用记者mRNA来评估翻译过程性.
主要成果:
- 建立了一个功能混合翻译系统.
- 该系统允许通过修改或未修改的核糖体来研究翻译.
- 可以有效地评估翻译延长过程性.
结论:
- 开发的混合翻译系统克服了标准RRL的局限性.
- 该协议促进了对核糖体功能和转化控制的研究.
- 该系统适用于研究异质核糖体的性能.
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