完整的蛋白质条形码可以一次性识别CRISPRi菌株及其代谢状态
Vanessa Pahl1, Paul Lubrano2, Felicia Troßmann2
1Interfaculty Institute of Microbiology and Infection Medicine, University of Tübingen, Auf der Morgenstelle 24, 72076 Tübingen, Germany; Cluster of Excellence "Controlling Microbes to Fight Infections", University of Tübingen, 72076 Tübingen, Germany.
Cell reports methods
|November 27, 2024
概括
完整的蛋白质条形码使用质谱法同时检测工程细菌中的遗传条形码和代谢物. 这种方法可以加速对大型细菌库进行选,用于研究和开发.
科学领域:
- 生物技术是生物技术.
- 分析化学 分析化学
- 代谢学 代谢学 代谢学
背景情况:
- 对基因工程细菌库的选对研究至关重要.
- 工程图书馆的菌株特定检测通常依赖于基于DNA的条形码.
- 质谱仪为分子检测提供了高灵敏度.
研究的目的:
- 引入完整的蛋白质条形码,以便同时测量蛋白质条形码和代谢物.
- 为了证明流量注射质谱法 (FI-MS) 对此应用的实用性.
- 评估图书馆选方法的可扩展性和适用性.
主要方法:
- 开发了一种蛋白质条形码系统,使用具有N端标签的ubiquitin.
- 采用流注射质谱法 (FI-MS) 来同时检测完整的乌比奎丁蛋白质和代谢物.
- 构建和分析了六种针对代谢酶的全方位素条形编码CRISPR干扰 (CRISPRi) 菌株.
主要成果:
- FI-MS成功检测到完整的泛素蛋白,并确定了N端条码质量.
- 主要代谢物的相对度与条码检测同时进行测量.
- 在CRISPRi向的途径中观察到明显的代谢组变化,与检测到的条形码相关.
- 通过测量132个在微型板中的泛素条形码来证明可扩展性.
结论:
- 完整的蛋白质条形码可以快速同时检测图书馆条形码和细胞内代谢物.
- 这种方法扩大了生物研究中基于质谱的条形码的可能性.
- 该技术可促进工程细菌库的高通量选.
关键词:
CP: 生物技术 生物技术CP:新陈代谢过程中的新陈代谢.这就是CRISPR干扰的原因.条码编码 条码编码质谱测量质谱测量质谱测量质量测量质谱测量质量测量质量测量质量测量质量测量质量测量质量测量质量测量质量测量质量测量质量测量质量测量质量测量质量测量质量测量质量测量质量测量质量测量质量测量质量测量质量测量代谢生物组的代谢生物组顶向下的蛋白质组学更多相关视频
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