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Understanding the Impact of Temperate Bacteriophages on Their Lysogens Through Transcriptomics
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Understanding the Impact of Temperate Bacteriophages on Their Lysogens Through Transcriptomics

Understanding the Impact of Temperate Bacteriophages on Their Lysogens Through Transcriptomics

DOI:
10.3791/64945-v

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09:23 min

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January 05, 2024

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, , , , , ,
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

Chapters

  • 00:04Introduction
  • 01:20Temporal Direct Enumeration of Spontaneous Induction
  • 02:56Preparation of Un‐Induced and Induced Lysogen Cultures for RNA Extraction
  • 05:00Standard Curve and Quantitative‐PCR
  • 06:42Results: Understanding the Impact of Prophages on Their Hosts Using PFU and RT‐qPCR
  • 08:23Conclusion

Summary

Automatic Translation

Automatic Translation

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.

Tags

Keywords: Temperate BacteriophagesProphagesTranscriptomic ApproachBacterial HostsLysogenLytic CycleGene ExpressionBacterial PathogensTherapeutic TargetsRNA IntegrityIndicator HostLysogensInfective Phage ParticlesRifampicin ResistantTop Agar Layer

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