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Related Experiment Videos

A synthetic phage lambda regulatory circuit.

Shota Atsumi1, John W Little

  • 1Department of Biochemistry, University of Arizona, Tucson, AZ 85721, USA.

Proceedings of the National Academy of Sciences of the United States of America
|December 1, 2006
PubMed
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Researchers engineered phage lambda by replacing key genetic components. They discovered an unexpected role for Lac repressor in maintaining lysogeny, revealing a novel regulatory network in bacteriophage lambda.

Area of Science:

  • Molecular Biology
  • Synthetic Biology
  • Genetics

Background:

  • Bacteriophage lambda's gene regulatory circuit controls its life cycle (lytic vs. lysogenic).
  • Modifying natural circuits offers a way to study and evolve their functions.
  • Previous work replaced lambda Cro with a tunable Lac repressor module.

Purpose of the Study:

  • To replace lambda CI repressor with a tunable Tet repressor module.
  • To investigate the resulting phage lambda variants' behavior and regulatory properties.
  • To explore the potential for engineering novel gene regulatory circuits.

Main Methods:

  • Genetic engineering of bacteriophage lambda.
  • Replacement of the CI repressor with a Tet repressor module.

Related Experiment Videos

  • Combinatorial selection to isolate functional phage variants.
  • Analysis of lysogen stability and induction responses.

    • Main Results:

    • Isolated phage variants exhibited lytic growth and stable lysogeny.
    • Tet repressor ligand addition induced prophage, as expected.
    • Unexpectedly, Lac repressor ligand addition also induced lysogens, indicating its essential role.
    • This suggests an altered, more complex regulatory wiring diagram than wild-type lambda.

    Conclusions:

    • The engineered phage lambda exhibits a novel regulatory mechanism for lysogeny involving both Tet and Lac repressors.
    • This complexity may compensate for the absence of wild-type CI repressor features.
    • The methodology is broadly applicable for creating custom, small-molecule-regulated gene circuits.