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Related Experiment Videos

Filamentous phage assembly: morphogenetically defective mutants that do not kill the host.

G P Smith1

  • 1Division of Biological Sciences, University of Missouri, Columbia 65211.

Virology
|November 1, 1988
PubMed
Summary

Filamentous phage assembly can be studied without host death using a modified phage (fd-tet). This system reveals essential genes for phage assembly and clarifies the role of a cis-acting sequence in phage production.

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Area of Science:

  • Molecular Biology
  • Virology
  • Bacteriophage Genetics

Background:

  • Filamentous phage assembly occurs via extrusion without host lysis.
  • Blocked assembly leads to host death due to accumulated phage DNA and proteins.
  • Phage fd-tet, with a tetracycline-resistance gene, exhibits reduced host toxicity.

Purpose of the Study:

  • To investigate filamentous phage assembly mechanisms.
  • To identify genes essential for virion formation and maturation.
  • To analyze the function of the cis-acting sequence 'mos' in phage yield.

Main Methods:

  • Utilized phage fd-tet for propagation in tetracycline-containing media, minimizing host morbidity.
  • Employed transfection to initiate cultures, enabling sensitive assays of low-level phage production.

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  • Studied morphogenetically defective mutants to determine gene requirements for assembly.

    • Main Results:

    • Genes III, VI, I, and IV are not essential for the initial DNA-protein complex formation.
    • Genes I and/or IV are critically required for the subsequent assembly process.
    • The cis-acting sequence 'mos' demonstrated variable effects on phage yield depending on experimental conditions.

    Conclusions:

    • Established a system to study phage assembly independent of host cell death.
    • Elucidated the roles of specific genes in the filamentous phage life cycle.
    • Provided new insights into the regulation of phage production and assembly efficiency.