建立大肠杆菌BW2511噬菌体-宿主相互作用的定性趋化敏感试验

The Meducator Pub Date : 2019-11-26 DOI:10.35493/medu.36.22
Aaron Wen, Esra Rakab
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引用次数: 0

摘要

为了更好地理解噬菌体和宿主细菌之间的相互作用,需要观察的一个因素是趋化性。趋化性描述了细菌偏向或远离某种物质的定向运动。这种现象是由翻滚事件的抑制或增强造成的,这些事件每隔几秒钟就会随机化宿主的方向。化学引诱剂和化学驱避剂的存在可以改变发生的翻滚事件的数量。为了阐明噬菌体在细菌趋化性中的作用,本项目旨在优化一种定性观察大肠杆菌BW2511 (WT)趋化性的方法,并与cheA缺失突变株进行比较。试验了三种主要的检测方法:Tris HCl平板法、Tris HCl微毛细管法和TTC Eppendorf法。在所有测试的方法中,只有TTC Eppendorf法与TTC指示剂一致显示趋化性。本研究继续使用Eppendorf实验,并确定各种噬菌体是否可以吸引大肠杆菌BW2511,使其在一层含0.4% pbs的琼脂上化学移动,并抵抗重力。
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Developing a qualitative chemotaxis-sensitive assay for bacteriophage-host interactions on E. coli BW2511
To better understand the interactions between bacteriophages and host bacteria, one factor to observe is chemotaxis. Chemotaxis describes the biased directional movement of bacteria either towards or away from a substance. This phenomenon is created by the inhibition or potentiation of tumbling events, which randomize the direction of the host every few seconds. The presence of chemoattractants and chemorepellents can modify the number of tumbling events that occur. In order to elucidate the role of bacteriophages in bacterial chemotaxis, this project aimed to optimize an assay to qualitatively observe chemotaxis within E. coli BW2511 (WT) in comparison to a mutant strain with cheA deletion. Three main assay methods were tested: Tris HCl plate assays, Tris HCl microcapillary assays, and TTC Eppendorf assays. Of all the methods tested, only TTC Eppendorf assays with TTC indicator consistently showed chemotaxis. This study continued to use the Eppendorf assay and determined if various bacteriophages could attract E. coli BW2511 to chemotactically move up a layer of 0.4% PBS-based agar and against gravitational pull.
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