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Related Experiment Videos

Isolation of lambda transducing phage with the bio genes inserted between lambda genes P and Q.

N W Charon, A M Campbell, S C Stamm

    Genetics
    |May 1, 1980
    PubMed
    Summary

    Researchers constructed biotin-transducing phages by inserting bio genes into bacteriophage lambda for gene fusion experiments. These phages showed instability, losing bio genes during propagation, suggesting issues with lambda Q gene fusion for successful phage propagation.

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    Area of Science:

    • Molecular Biology
    • Bacteriophage Genetics
    • Genetic Engineering

    Background:

    • Bacteriophage lambda is a well-characterized model organism for genetic studies.
    • The bio operon, involved in biotin synthesis, is a target for genetic manipulation.
    • Gene fusion strategies are employed to study gene regulation and protein interactions.

    Purpose of the Study:

    • To construct plaque-forming, biotin-transducing bacteriophage lambda with bio genes inserted for future lambda Q gene fusion.
    • To investigate the feasibility of fusing the lambda Q gene to the bio operon.
    • To characterize the resulting recombinant phages and assess their stability and propagation.

    Main Methods:

    • Construction of a defective temperature-sensitive lysogen containing bio genes adjacent to lambda genes.

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  • Screening for heat-resistant survivors with specific deletions within the bio operon and phage lambda.
  • Isolation and characterization of plaque-forming, biotin-transducing phages.
  • Analysis of gene order, bio gene content, and phage gene duplication.
  • Assessment of phage propagation under specific conditions.

    • Main Results:

    • Five heat-resistant survivors with desired deletion endpoints were identified.
    • Two strains yielded plaque-forming, biotin-transducing phages upon induction.
    • Isolated phages exhibited instability, losing bio genes during propagation due to partial phage gene duplication.
    • One phage, lambda bioq1b221, showed a duplication not involving the entire lambda Q gene.
    • A recombinant phage failed to plaque under biotin-derepression conditions.

    Conclusions:

    • The constructed phages are unstable, losing bio genes upon propagation, indicating challenges with partial phage gene duplication.
    • Insufficient lambda Q gene product is likely synthesized when fused to the bio operon, hindering phage propagation.
    • Further optimization is needed for successful lambda Q gene fusion to the bio operon for functional gene expression.