一个连续的无细胞翻译系统,能够生产高产量的多
A S Spirin1, V I Baranov, L A Ryabova
1Institute of Protein Research, Academy of Sciences, Moscow Region, USSR.
概括
这项研究开发了一种持续的无细胞翻译系统,用于持续的蛋白质合成. 无论是 prokaryotic 和 eukaryotic 系统都证明了长期的,高速的多产量超过 20 小时.
科学领域:
- 生物化学 生物化学
- 分子生物学分子生物学
- 合成生物学 合成生物学
背景情况:
- 无细胞蛋白合成 (CFPS) 系统为生产蛋白质提供了一个多功能平台.
- 传统的批量CFPS系统受到产品抑制和基质耗尽的限制.
- 连续流系统的目标是克服这些长时间合成的局限性.
研究的目的:
- 开发和描述一个连续流动的无细胞翻译系统.
- 评估 prokaryotic 和 eukaryotic CFPS 的长期稳定性和效率.
- 评估系统生产特定蛋白质产品的能力.
主要方法:
- 构建了一个连续流动的无细胞翻译系统,具有料缓冲区和产品清除.
- 测试了原生生物 (大肠杆菌) 和真核生物 (小麦芽) 无细胞系统.
- 使用MS2菌体RNA,体马赛克病毒RNA4和合成酸mRNA作为模板.
主要成果:
- 连续系统维持了高,恒定的多合成速率数小时.
- Prokaryotic 和 eukaryotic 系统在 20 小时内产生了每 RNA 模板的 ~ 100 个病毒外套蛋白质副本.
- 这两种系统在40个小时内产生了每个mRNA模板的150-300个卡尔西托尼多副本.
结论:
- 连续流动的无细胞翻译系统可实现长时间和高效的蛋白质合成.
- 这些系统在 prokaryotic 和 eukaryotic 环境中都表现出强大的性能.
- 开发的系统适合持续生产各种多,包括病毒蛋白和治疗性.
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