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通过丝裂霉素C介导的诱导结合流式细胞术分析检测原噬菌体

Detecting Prophages by Mitomycin C-Mediated Induction Coupled to Flow Cytometry Analysis.

作者信息

Oliveira Joana, Mahony Jennifer, Hanemaaijer Laurens, Kouwen Thijs R H M, Neve Horst, MacSharry John, van Sinderen Douwe

机构信息

School of Microbiology, University College CorkCork, Ireland.

APC Microbiome Institute, University College CorkCork, Ireland.

出版信息

Front Microbiol. 2017 Jul 19;8:1343. doi: 10.3389/fmicb.2017.01343. eCollection 2017.

Abstract

Most analyzed strains are predicted to harbor one or more prophage genomes within their chromosome; however, the true extent of the inducibility and functionality of such prophages cannot easily be deduced from sequence analysis alone. Chemical treatment of lysogenic strains with Mitomycin C is known to cause induction of temperate phages, though it is not always easy to clearly identify a lysogenic strain or to measure the number of released phage particles. Here, we report the application of flow cytometry as a reliable tool for the detection and enumeration of released lactococcal prophages using the green dye SYTO-9.

摘要

大多数分析菌株预计在其染色体中含有一个或多个前噬菌体基因组;然而,仅从序列分析中不容易推断出此类前噬菌体的可诱导性和功能的真实程度。已知用丝裂霉素C对溶原性菌株进行化学处理会导致温和噬菌体的诱导,尽管要清楚地鉴定溶原性菌株或测量释放的噬菌体颗粒数量并不总是那么容易。在这里,我们报告了流式细胞术作为一种可靠工具的应用,该工具可使用绿色染料SYTO-9检测和计数释放的乳球菌前噬菌体。

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