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通过高通量毛细管检测法对大肠杆菌趋化反应的优化。

Fine-tuning of chemotactic response in E. coli determined by high-throughput capillary assay.

机构信息

The Department of Physics, The James Franck Institute, The Institute for Biophysical Dynamics, University of Chicago, Chicago, IL 60637, USA.

出版信息

Curr Microbiol. 2011 Mar;62(3):764-9. doi: 10.1007/s00284-010-9778-z. Epub 2010 Oct 24.

Abstract

In E. coli, chemotactic behavior exhibits perfect adaptation that is robust to changes in the intracellular concentration of the chemotactic proteins, such as CheR and CheB. However, the robustness of the perfect adaptation does not explicitly imply a robust chemotactic response. Previous studies on the robustness of the chemotactic response relied on swarming assays, which can be confounded by processes besides chemotaxis, such as cellular growth and depletion of nutrients. Here, using a high-throughput capillary assay that eliminates the effects of growth, we experimentally studied how the chemotactic response depends on the relative concentration of the chemotactic proteins. We simultaneously measured both the chemotactic response of E. coli cells to L: -aspartate and the concentrations of YFP-CheR and CheB-CFP fusion proteins. We found that the chemotactic response is fine-tuned to a specific ratio of [CheR]/[CheB] with a maximum response comparable to the chemotactic response of wild-type behavior. In contrast to adaptation in chemotaxis, that is robust and exact, capillary assays revealed that the chemotactic response in swimming bacteria is fined-tuned to wild-type level of the [CheR]/[CheB] ratio.

摘要

在大肠杆菌中,趋化行为表现出完美的适应性,对趋化蛋白(如 CheR 和 CheB)的细胞内浓度变化具有很强的鲁棒性。然而,完美适应的鲁棒性并不明确意味着趋化反应具有鲁棒性。以前关于趋化反应鲁棒性的研究依赖于群体游动测定,该测定可能受到除趋化作用以外的过程的干扰,如细胞生长和营养物质的消耗。在这里,我们使用一种消除生长影响的高通量毛细管测定法,实验研究了趋化反应如何取决于趋化蛋白的相对浓度。我们同时测量了大肠杆菌细胞对 L:-天冬氨酸的趋化反应和 YFP-CheR 和 CheB-CFP 融合蛋白的浓度。我们发现,趋化反应被精细地调谐到 CheR/CheB 的特定比例,最大响应可与野生型行为的趋化响应相媲美。与适应趋化作用的鲁棒性和精确性不同,毛细管测定法表明,游动细菌的趋化反应被精细地调谐到 CheR/CheB 比值的野生型水平。

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