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Inducer-dependent conditional-lethal mutant animal viruses.

Y F Zhang1, B Moss

  • 1Laboratory of Viral Diseases, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, MD 20892.

Proceedings of the National Academy of Sciences of the United States of America
|February 15, 1991
PubMed
Summary
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Researchers engineered a conditional-lethal vaccinia virus using Escherichia coli lac operon elements. This novel virus requires an inducer for replication, offering a new tool for virology research and control.

Area of Science:

  • Virology
  • Molecular Biology
  • Genetic Engineering

Background:

  • Conditional-lethal viruses are valuable tools for studying viral replication and developing antiviral strategies.
  • The Escherichia coli lac operon provides a well-characterized system for inducible gene expression.

Purpose of the Study:

  • To construct an animal virus with an inducer-dependent conditional-lethal phenotype.
  • To utilize the regulatory elements of the lac operon for precise control of viral gene expression.

Main Methods:

  • Integration of the lac operon repressor gene into the vaccinia virus genome for constitutive expression.
  • Insertion of the lac operator adjacent to the promoter of a vaccinia virus gene encoding an 11-kDa protein.
  • Culturing the engineered virus in the presence (permissive) or absence (non-permissive) of the inducer.

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Main Results:

  • The engineered vaccinia virus exhibited conditional lethality, requiring the inducer for plaque formation and high viral yield.
  • Viral growth was significantly reduced (over 1000-fold) under non-permissive conditions.
  • Transcription of the targeted 11-kDa protein gene was strictly dependent on the inducer.

Conclusions:

  • The lac operon system can be effectively employed to create inducer-dependent conditional-lethal mutant viruses.
  • This strategy provides a robust method for controlling viral replication and offers potential applications in other viral systems.