建立基于人工 ribozyme 开关的对托布拉米辛敏感的全细胞微生物传感器
Zhenzhen Wang1, Jiawen Cheng1, Guimin Dai1
1School of Life Science, Anhui Medical University, Hefei 230032, China.
Life (Basel, Switzerland)
|July 29, 2023
概括
研究人员开发了一种新型的托布拉米辛生物传感器,使用头 ribozyme (HHR) 和aptamer. 这种全细胞传感器允许在食品和尿液样本中快速,灵敏地检测托布拉米辛.
科学领域:
- 分子生物学分子生物学
- 生物技术是生物技术.
- 生物化学 生物化学
背景情况:
- 食品和环境样本中的抗生素残留物构成重大风险.
- 开发敏感和快速检测方法对抗生素如托布拉米至关重要.
- 现有的生物传感器通常需要复杂的样品准备或恶劣的条件.
研究的目的:
- 构建和验证一种全细胞微生物传感器,用于托布拉米检测.
- 利用体内查和计算分析来优化生物传感器结构.
- 为开发基于人工 ribozyme 的生物传感器制定一个普遍适用的策略.
主要方法:
- 托布拉米辛的阿普坦与来自 * Schistosoma mansoni * 的头 ribozyme (HHR) 的融合,以创建依赖于托布拉米辛的全细胞微生物感应器 (tob-HHAz).
- 将模块集成到单个RNA序列中,以便轻松引入大肠杆菌.
- 高通量体内查和体内验证对托布拉米敏感RNA结构.
- 对RNA结构的计算预测,以分析形状和检测机制.
主要成果:
- 鉴定了三种对托布拉米辛敏感的RNA结构,并验证了它们的自我分裂活性.
- 两个传感器表现出对添加牛奶的高响应性,检测极限约为40nm.
- 一个传感器显示线性检测范围从30到650nM,在的尿液中检测的30nM极限,表现出良好的选择性.
结论:
- 开发的托布拉米生物传感器 (tob-HHAz) 提供了对抗生素残留物的敏感和快速检测.
- 在体内查和计算分析策略的结合有效地优化了生物传感器结构.
- 这种方法为开发基于人工 ribozyme 的生物传感器提供了一个多功能平台,用于检测活细胞中的各种物质.
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