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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
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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.

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.