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Detecting Lactococcus lactis Prophages by Mitomycin C-Mediated Induction Coupled to Flow Cytometry Analysis.
Joana Oliveira1, Jennifer Mahony1,2, Laurens Hanemaaijer3
1School of Microbiology, University College CorkCork, Ireland.
Frontiers in Microbiology
|August 4, 2017
Summary
Flow cytometry offers a reliable method for detecting and counting lactococcal prophages released from bacteria. This technique uses SYTO-9 dye for accurate enumeration of these essential viral elements.
Area of Science:
- Microbiology
- Virology
- Bacteriology
Background:
- Lactococcus lactis strains often contain prophage genomes, but their activity is hard to determine from DNA sequences alone.
- Inducing prophages with chemicals like Mitomycin C is common, but identifying lysogenic strains and quantifying phage release is challenging.
Purpose of the Study:
- To establish flow cytometry as a dependable method for detecting and enumerating prophages released from Lactococcus lactis.
- To utilize SYTO-9 fluorescent dye for accurate prophage quantification.
Main Methods:
- Applying flow cytometry for the detection of released lactococcal prophages.
- Using the green fluorescent dye SYTO-9 to label and quantify phage particles.
Main Results:
- Flow cytometry provides a reliable means to detect prophages released from Lactococcus lactis.
- SYTO-9 staining enables accurate enumeration of these released phage particles.
Conclusions:
- Flow cytometry is an effective tool for studying prophage induction and release in Lactococcus lactis.
- This method enhances the ability to assess prophage functionality beyond sequence analysis.
