A palette of fluorophores that are differentially accumulated by wild-type and mutant strains of Escherichia coli: surrogate ligands for profiling bacterial membrane transporters.
A palette of fluorophores that are differentially accumulated by wild-type and mutant strains of Escherichia coli: surrogate ligands for profiling bacterial membrane transporters.
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由野生型和突变型大肠杆菌菌株不同积累的荧光团的调色板：分析细菌膜转运体的替代配体。
DOI：
10.1099/mic.0.001016
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发表时间：
2021-03
期刊：
影响因子：
--
通讯作者：
Kell DB
中科院分区：
文献类型：
--
作者：
Salcedo-Sora JE;Jindal S;O'Hagan S;Kell DB
我们以前的工作表明，两种常用的荧光染料，积累了野生型大肠杆菌MG 1655的差异在单基因敲除菌株的运输，他们也可能被用作替代品在流式细胞仪转运蛋白测定。我们总结了这些染料的理想性能，并在这里调查143个候选染料。我们最终将它们分类（基于信号，积累水平和成本）到39个市售和负担得起的荧光团的调色板中，这些荧光团通过“Keio”菌株BW 25113的野生型细胞显著积累，如流式细胞术测量的。化学信息学分析表明它们的相似性和它们（更可观的）结构差异。我们描述了pH值和外排泵抑制剂氯丙嗪对染料积累的影响。即使是“野生型”MG 1655和BW 25113菌株在吸收此类染料的能力上也可能存在显著差异。我们说明了我们的染料的高度差异摄取到具有特定病变或过表达水平的三种特定转运蛋白或转运蛋白组分（yhjV，yihN和tolC）的菌株中。所描述的染料的相对小的集合提供了一种快速、廉价、方便和信息丰富的方法来评估微生物生理学和膜转运蛋白功能的表型。
Our previous work demonstrated that two commonly used fluorescent dyes that were accumulated by wild-type Escherichia coli MG1655 were differentially transported in single-gene knockout strains, and also that they might be used as surrogates in flow cytometric transporter assays. We summarize the desirable properties of such stains, and here survey 143 candidate dyes. We eventually triage them (on the basis of signal, accumulation levels and cost) to a palette of 39 commercially available and affordable fluorophores that are accumulated significantly by wild-type cells of the ‘Keio’ strain BW25113, as measured flow cytometrically. Cheminformatic analyses indicate both their similarities and their (much more considerable) structural differences. We describe the effects of pH and of the efflux pump inhibitor chlorpromazine on the accumulation of the dyes. Even the ‘wild-type’ MG1655 and BW25113 strains can differ significantly in their ability to take up such dyes. We illustrate the highly differential uptake of our dyes into strains with particular lesions in, or overexpressed levels of, three particular transporters or transporter components (yhjV, yihN and tolC). The relatively small collection of dyes described offers a rapid, inexpensive, convenient and informative approach to the assessment of microbial physiology and phenotyping of membrane transporter function.
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影响因子：
8.8
作者：
Cardinale, Stefano;Tueros, Felipe Gonzalo;Sommer, Morten Otto Alexander
通讯作者：
Sommer, Morten Otto Alexander
影响因子：
2.4
作者：
Currin, Andrew;Swainston, Neil;Kell, Douglas B.
通讯作者：
Kell, Douglas B.
大肠杆菌K-12的构造框架，单基因敲除突变体：Keio Collection。
影响因子：
9.9
作者：
通讯作者：
--
影响因子：
5.2
作者：
Baugh, Stephanie;Phillips, Charlotte R.;Webber, Mark A.
通讯作者：
Webber, Mark A.
影响因子：
3.2
作者：
DEZWAIG, RN;LURIA, SE
通讯作者：
LURIA, SE