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Related Experiment Videos

Specificity in DNA recognition by phage integrases.

Allan Campbell1, Alice del-Campillo-Campbell, Michael L Ginsberg

  • 1Department of Biological Sciences, Stanford University, Stanford, CA 94305, USA. amc@stanford.edu

Gene
|December 7, 2002
PubMed
Summary

Lambdoid coliphages exhibit functional polymorphism due to natural recombination. Selection favors new site recognition specificities, as demonstrated by differing integration behaviors of phages 21 and e14.

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Area of Science:

  • Microbiology
  • Molecular Biology
  • Genetics

Background:

  • Lambda-related (lambdoid) coliphages are bacteriophages that infect E. coli.
  • These phages undergo frequent natural recombination, leading to functional polymorphism.
  • The integration module of these phages is known to exhibit polymorphism.

Purpose of the Study:

  • To investigate the role of selection in driving the polymorphism of phage integration modules.
  • To understand the molecular basis of site recognition specificity in lambdoid phages.
  • To compare the integration and excision behaviors of related phages and elements.

Main Methods:

  • Analysis of amino acid changes affecting phage site recognition specificity.
  • In vitro assays to study the integration and excision of phage 21 and element e14.

Related Experiment Videos

  • Oligonucleotide inhibition assays to define sequence specificity.

    • Main Results:

    • Polymorphism in the integration module likely results from selection for new site recognition specificities.
    • Phages lambda and HK022 integration specificity can be altered by changes in five noncontiguous amino acids.
    • Phage 21 and element e14, though integrating at the same site, exhibit distinct core and arm site recognition specificities.
    • In vitro assays confirmed these differing specificities for e14 and 21.
    • ds arm site oligonucleotides precisely defined the sequence specificity.

    Conclusions:

    • Natural recombination and selection contribute to functional polymorphism in lambdoid coliphages.
    • Selection appears to favor novel specificities rather than improvements in existing ones.
    • Specific amino acid changes can significantly alter phage integration site recognition.
    • Phage 21 and element e14 represent distinct examples of site recognition specificity within the same integration locus.