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Related Experiment Videos

On the clustered exchanges of the RecBCD pathway operating on phage lambda

F W Stahl1, C E Shurvinton, L C Thomason

  • 1Institute of Molecular Biology, University of Oregon, Eugene 97403-1229, USA.

Genetics
|March 1, 1995
PubMed
Summary

Recombination between phage lambda and plasmids in E. coli reveals that plasmid homology influences phage-phage exchanges. This relationship is independent of the Rap function, suggesting distinct mechanisms for different recombination events.

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Area of Science:

  • Molecular Biology
  • Microbiology
  • Genetics

Background:

  • Phage lambda recombination in Escherichia coli is a complex process involving host and phage factors.
  • Previous studies indicated that recombination with a plasmid can alter the outcome of phage lambda x phage lambda exchanges.

Purpose of the Study:

  • To investigate the mechanistic relationship between lambda x plasmid and lambda x lambda recombination events.
  • To determine if the homology between lambda and the plasmid is essential for the observed alterations in recombination outcomes.
  • To assess the role of lambda's Rap function in these recombination pathways.

Main Methods:

  • Conducted lytic cycle crosses of Red- Gam- phage lambda in rec+ Escherichia coli.
  • Utilized plasmids with homology to phage lambda.

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  • Analyzed lambda x lambda and lambda x plasmid recombinants formed via RecBCD-mediated recombination.
  • Examined the effect of removing plasmid homology on recombination patterns.

    • Main Results:

    • The relationship between lambda x plasmid and lambda x lambda exchanges remained consistent even when plasmid homology was removed.
    • This supports the model that reciprocal exchange (cointegrate formation) is mechanistically separable from nonreciprocal lambda x lambda exchange.
    • Lambda's Rap function modulated the ratio of cointegrate formation (splices) to marker pick-up (patches) in RecBCD-mediated lambda x plasmid recombination.

    Conclusions:

    • Plasmid homology influences phage lambda recombination outcomes, but is not essential for the observed relationship between different exchange types.
    • Reciprocal and nonreciprocal exchange mechanisms in phage lambda recombination are distinct.
    • The Rap function plays a role in regulating the balance between different outcomes in lambda x plasmid recombination.