微流体对电泳运动移动性的选 转移 阐明 рибо开关 结合功能
Kelly Karns1, Jacob M Vogan, Qian Qin
1San Francisco Graduate Program in Bioengineering, University of California, Berkeley, California 94720, USA.
Journal of the American Chemical Society
|January 25, 2013
概括
我们开发了一种快速的微流体移动性转移试验,用于量化 рибо开关-连接体结合和构造变化. 这种高通量工具可以加速发现新型的核糖突变器和潜在的抗生素点.
科学领域:
- 分子生物学分子生物学
- 生物物理学的生物物理.
- 生物化学 生物化学
背景情况:
- рибо开关是RNA传感器,通过改变结合小分子代谢物后的构造来调节基因表达.
- 现有的研究 рибо开关-连接体相互作用的方法往往耗时且缺乏高通量能力.
- 需要一个快速的,定量工具来选riboswitch及其连接体结合.
研究的目的:
- 引入和验证微流体移动性转移试验,用于高通量,对 рибо开关-连接体结合的定量分析.
- 为了证明该试验能够快速选和验证候选的核糖开关.
- 建立一个可扩展的平台,用于表征 рибо开关的结构变化和结合动力学.
主要方法:
- 开发一种微流体移动性转移试验,用于精确量化 рибо交换机-连接体结合.
- 从热友和冷友细菌中选和验证了五个候选SAM-I рибо开关.
- 使用微流体试验量化平衡分离常数 (K(d)) 并与线式探测结果进行比较.
主要成果:
- 微流体检测屏幕在传统基板检测所需时间的0.3% (3.2分钟与1020分钟相比) 中进行 рибо切换.
- 该测试提供了增强的分辨率,量化和可重复性,用于检测小的移动性转移和形状变化.
- 对于已知和候选SAM-I рибо开关,均准确量化了平衡分离常数 (K(d)).
结论:
- 微流体移动性转移试验是一种快速,灵敏和可扩展的工具,用于研究 рибо开关-连接体相互作用.
- 这项技术加速了新型 рибо开关的发现和选择.
- 该试验有助于开发新的抗生素,这些抗生素针对细菌的核糖转换器.
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相关概念视频
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