基于RNA的光生物传感器用于第二信使循环di-AMP的活细胞成像
Colleen A Kellenberger1, Chen Chen, Aaron T Whiteley2
1†Department of Chemistry, University of California, Berkeley, California 94720, United States.
Journal of the American Chemical Society
|May 13, 2015
概括
科学家们开发了光生物传感器,以可视化细菌中的循环二AMP (cdiA). 这项创新有助于了解细菌代谢和抗生素耐药性,验证病原体和古生物中的酶.
科学领域:
- 微生物学 微生物学
- 分子生物学分子生物学
- 生物化学 生物化学
背景情况:
- 循环二-AMP (cdiA) 是细菌中关键的第二信使,影响新陈代谢和细胞壁完整性.
- 在Listeria monocytogenes中降低cdiA水平与对抗生素敏感性增加相关.
- 在各种细菌物种和古生物中,cdiA信号通路被保存.
研究的目的:
- 开发新的光生物传感器,用于可视化活细菌中的细胞内cdiA水平.
- 为了表征化酶PdeA和选二核酸环酶活性.
- 为了验证Clostridium difficile和Archaea中DisA样蛋白的功能.
主要方法:
- 菜2的阿普坦与cdiA的riboswitch连接体结合域的融合产生光生物传感器.
- 在活体中对活体Listeria monocytogenes中细胞内cdiA度的体内成像.
- 开发一种流细胞测量试验,用于选二核酸环酶活性.
主要成果:
- 成功生成并使用光生物传感器进行实时cdiA成像.
- 确定了二酶PdeA的催化域.
- 证实了来自Clostridium difficile和Methanocaldococcus jannaschii的DisA样蛋白的酶活性.
结论:
- 基于RNA的生物传感器能够有效地在活体中对Gram阳性细菌中的代谢物成像.
- 这项研究验证了第一个古老的二核酸环酶,扩大了这些酶的已知谱.
- 开发的生物传感器为研究cdiA信号和选不同微生物领域的酶活动提供了强大的工具.
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