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通过转录组学了解温带噬菌体对其溶原的影响

Journal JoVE
Biologie

This content is Free Access.

Journal JoVE Biologie

Understanding the Impact of Temperate Bacteriophages on Their Lysogens Through Transcriptomics

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通过转录组学了解温带噬菌体对其溶原的影响

DOI:

10.3791/64945-v

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09:23 min

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January 05, 2024

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Revathy Krishnamurthi, Enrique González-Tortuero, Grace Plahe, Ian B. Goodhead, Joanne L. Fothergill, Chloë E. James, Heather E. Allison

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

Chapitres

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

Traduction automatique

该协议能够揭示噬菌体对其宿主的影响。细菌培养物使用最能支持溶原状态的条件进行同步，从而限制了自发诱导。RT-qPCR明确区分噬菌体限制性基因和与噬菌体对照未偶联的基因与裂解复制周期中表达的基因。

Tags

Temperate Bacteriophages Prophages Transcriptomic Approach Bacterial Hosts Lysogen Lytic Cycle Gene Expression Bacterial Pathogens Therapeutic Targets RNA Integrity Indicator Host Lysogens Infective Phage Particles Rifampicin Resistant Top Agar Layer

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