التفاصيل البيبلوغرافية
| العنوان: |
Understanding the Impact of Temperate Bacteriophages on Their Lysogens Through Transcriptomics. |
| المؤلفون: |
Krishnamurthi R; Department of Clinical Infection, Microbiology and Immunology, Institute of Infection, Veterinary and Ecological Sciences (IVES), University of Liverpool., González-Tortuero E; School of Science, Engineering, and Environment, University of Salford., Plahe G; School of Science, Engineering, and Environment, University of Salford., Goodhead IB; School of Science, Engineering, and Environment, University of Salford., Fothergill JL; Department of Clinical Infection, Microbiology and Immunology, Institute of Infection, Veterinary and Ecological Sciences (IVES), University of Liverpool., James CE; School of Science, Engineering, and Environment, University of Salford., Allison HE; Department of Clinical Infection, Microbiology and Immunology, Institute of Infection, Veterinary and Ecological Sciences (IVES), University of Liverpool; hallison@liverpool.ac.uk. |
| المصدر: |
Journal of visualized experiments : JoVE [J Vis Exp] 2024 Jan 05 (203). Date of Electronic Publication: 2024 Jan 05. |
| نوع المنشور: |
Journal Article; Video-Audio Media |
| اللغة: |
English |
| بيانات الدورية: |
Publisher: MYJoVE Corporation Country of Publication: United States NLM ID: 101313252 Publication Model: Electronic Cited Medium: Internet ISSN: 1940-087X (Electronic) Linking ISSN: 1940087X NLM ISO Abbreviation: J Vis Exp Subsets: MEDLINE |
| أسماء مطبوعة: |
Original Publication: [Boston, Mass. : MYJoVE Corporation, 2006]- |
| مواضيع طبية MeSH: |
Bacteriophages*/genetics, Gene Expression Profiling ; Bacterial Typing Techniques ; Cell Count ; Chromosomes, Bacterial |
| مستخلص: |
Temperate phages are found integrated as prophages in the majority of bacterial genomes. Some prophages are cryptic and fixed in the bacterial chromosome, but others are active and can be triggered into a replicative form either spontaneously or by exposure to inducing factors. Prophages are commonly associated with the ability to confer toxin production or other virulence-associated traits on their host cell. More recent studies have shown they can play a much bigger role in altering the physiology of their hosts. The technique described here has enabled us to investigate how prophages affect gene expression in the opportunistic bacterium Pseudomonas aeruginosa. In this work, the growth of the wild-type P. aeruginosa strain PAO1 was compared with that of isogenic lysogens carrying different combinations of prophages from the Liverpool Epidemic Strain (LES) LESB58. In a lysogen culture, a proportion of bacterial cells will be supporting lytic bacteriophage replication (spontaneous induction) with a high level of expression per cell of late phage genes, such as those associated with the assembly of phage particles, thus masking the low-level gene expression associated with lysogen-restricted gene expression. The impact of spontaneous induction can thus obscure prophage gene expression across a lysogen population. Growth profiling experiments were used to identify spontaneous induction, which was minimal during the early exponential growth phase. This study reports how to prepare sample cultures during the early exponential growth phase and how to set up adequate controls despite low cell numbers. These protocols ensure the reliable and reproducible comparison of wild-type and lysogenic bacteria under various conditions, thus improving the transcriptomic profiling of prophage genomes and aiding in the identification of previously unrecognized prophage functions. |
| تواريخ الأحداث: |
Date Created: 20240122 Date Completed: 20240123 Latest Revision: 20240123 |
| رمز التحديث: |
20240123 |
| DOI: |
10.3791/64945 |
| PMID: |
38251745 |
| قاعدة البيانات: |
MEDLINE |
