JoVE Journal

Biology

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Understanding the Impact of Temperate Bacteriophages on Their Lysogens Through Transcriptomics

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1.8K Views

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09:23 min

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January 5th, 2024

DOI :

10.3791/64945-v

January 5th, 2024

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Revathy Krishnamurthi¹, Enrique González-Tortuero², Grace Plahe², Ian B. Goodhead², Joanne L. Fothergill¹, Chloë E. James², Heather E. Allison¹

¹Department of Clinical Infection, Microbiology and Immunology, Institute of Infection, Veterinary and Ecological Sciences (IVES), University of Liverpool, ²School of Science, Engineering, and Environment, University of Salford

Podsumowanie

This protocol enables the impact of prophages on their hosts to be revealed. Bacterial cultures are synchronized using conditions that best support the lysogenic state, limiting spontaneous induction. RT-qPCR unequivocally distinguishes prophage-restricted genes and those uncoupled from phage control from those that are expressed during the lytic replication cycle.

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Temperate Bacteriophages

Transcriptomics

Bacterial Hosts

Gene Expression

Therapeutic Targets

Culture Conditions

Dilution Series

Viable Bacterial Cells

Infective Phage Particles

Rozdziały w tym wideo

0:04

Introduction

1:20

Temporal Direct Enumeration of Spontaneous Induction

2:56

Preparation of Un‐Induced and Induced Lysogen Cultures for RNA Extraction

5:00

Standard Curve and Quantitative‐PCR

6:42

Results: Understanding the Impact of Prophages on Their Hosts Using PFU and RT‐qPCR

8:23

Conclusion

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