Quantification of plasmid loss in Escherichia coli cells by use of flow cytometry
Quantification of plasmid loss in Escherichia coli cells by use of flow cytometry
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DOI：
10.1016/s0378-1097(04)00015-1
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发表时间：
2004-03-12
影响因子：
2.1
通讯作者：
Hansen, LH
中科院分区：
文献类型：
--
作者：
Bahl, ML;Sorensen, SJ;Hansen, LH
开发了一种研究大肠杆菌细胞中质粒稳定性的方法，该方法利用流式细胞术的高速分析特性。为了区分携带质粒的细胞和无质粒的细胞，使用质粒编码的 Lac 阻遏蛋白来调节宿主细胞中染色体插入的绿色荧光蛋白基因的表达。由于无质粒细胞的绿色荧光表型，流式细胞术分析能够检测和定量无质粒细胞。所报告的系统提供实时分析，并且细菌群体中质粒丢失的检测水平非常低。这对于未来细菌群落中质粒稳定性和群体选择的研究可能很有用。 (C) 2004 年欧洲微生物学会联合会。由 Elsevier B.V. 出版。保留所有权利。
A method was developed to study plasmid stability in Escherichia coli cells, which utilised the high speed analysis properties of flow cytometry. To discriminate between plasmid-harbouring cells and plasmid-free cells a plasmid-encoded Lac repressor protein was used to regulate the expression of a chromosomally inserted green fluorescent protein gene in the host cells. Flow cytometric analysis enabled detection and quantification of plasmid-free cells due to their green fluorescent phenotype. The reported system offers real-time analysis in combination with a very low detection level of plasmid loss in bacterial populations. This could be useful in future investigations of plasmid stability and population selection in bacterial communities. (C) 2004 Federation of European Microbiological Societies. Published by Elsevier B.V. All rights reserved.