在铁饥饿和无氧生长后,量化Yersinia伪结核病III型分泌系统活动
Mané Ohanyan1, Diana Hooker-Romero1, David Balderas1
1Department of Microbiology and Environmental Toxicology, University of California, Santa Cruz.
Journal of visualized experiments : JoVE
|June 17, 2024
概括
这项研究详细介绍了一种诱导Yersinia伪结核病中的铁饥饿的方法,使研究人员能够在与宿主相关的条件下研究类型III分泌系统 (T3SS) 进行抗菌药物开发.
科学领域:
- 微生物学 微生物学
- 细菌学 细菌学是一门学科.
- 病变的发生和发病.
背景情况:
- 第三类分泌系统 (T3SS) 是格拉姆阴性病原体的关键毒性因子.
- 铁和氧气对T3SS的调节对哺乳动物感染至关重要.
- 针对T3SS是一种有前途的抗微生物发现策略.
研究的目的:
- 描述一个强大的方法,以铁饥饿的Yersinia伪结核病.
- 评估氧气可用性对T3SS表达的影响.
- 在与宿主相关的条件下为研究T3SS活动提供框架.
主要方法:
- 通过酸洗玻璃器皿,合剂和长期培养在缺乏铁的介质中,对Y. pseudotuberculosis的铁饥饿.
- 在37°C的有氧和无氧条件下培养.
- 通过使用SDS-PAGE和银色染色分析分泌的效应蛋白来量化T3SS活性.
主要成果:
- 开发的方法有效地诱导铁饥饿,促进T3SS表达.
- 氧气的可用性会影响T3SS活动.
- 该协议允许对T3SS活动进行可访问的测量.
结论:
- 这种方法提供了一种可靠的方法来研究Y. pseudotuberculosis中的T3SS调节和活性.
- 该协议可以适应其他具有高效铁吸收系统的病原体.
- 这些发现支持开发针对T3SS的新型抗菌药物.
相关概念视频
Microbial Growth Measurement: Direct Methods
2.7K
Direct methods for measuring microbial populations in a culture are essential tools in microbiology, providing quantitative data for various applications. Among these, microscopic counts, plate counts, and serial dilution are widely used techniques, each with unique principles and applications.Microscopic CountsMicroscopic counting involves the use of a Petroff-Hausser chamber, a specialized microscope slide with a grid and defined depth. By observing a liquid culture under a microscope,...
2.7K
Microbial Growth Measurement: Indirect Methods
2.4K
Estimating microbial growth is essential for understanding population dynamics and environmental adaptations. Indirect methods provide valuable insights by measuring parameters such as turbidity, metabolic activity, and biomass, enabling efficient and reproducible assessments.During exponential growth, microbial cells scatter light proportionally to their biomass, a principle used in turbidity measurements. About one million cells per milliliter produce detectable scattering, which a...
2.4K
Stringent Response in E. coli
477
Bacterial growth is closely tied to nutrient availability, with cells proliferating exponentially under favorable conditions and entering a stationary phase when resources become scarce. This transition is mediated by a regulatory mechanism known as the stringent response, which allows bacteria to adapt to nutrient deprivation by modulating gene expression and metabolic activity.During nutrient scarcity, intracellular amino acid levels decline. It results in the accumulation of uncharged tRNAs...
477


