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Resolvase-like serine recombinase mediates integration/excision in the bacteriophage φRSM

Ahmed Askora1, Takeru Kawasaki, Makoto Fujie

  • 1Department of Molecular Biotechnology, Graduate School of Advanced Sciences of Matter, Hiroshima University, Higashi-Hiroshima 739-8530, Japan.

Journal of Bioscience and Bioengineering
|November 2, 2010
PubMed

Insights

This study shows that ORF14 from the filamentous phage ϕRSM1 acts as a serine recombinase, mediating integration and excision for phage DNA in Ralstonia solanacearum. This is the first evidence of such a mechanism in filamentous phages.

Area of Science:

  • Microbiology
  • Molecular Biology
  • Genetics

Background:

  • Filamentous phages are important genetic elements in bacteria.
  • Ralstonia solanacearum is a plant pathogen with associated prophages.
  • Site-specific recombinases play crucial roles in phage lifecycle.

Purpose of the Study:

  • To characterize the integrative and excisive recombination functions of ORF14 from the filamentous phage ϕRSM1.
  • To investigate the role of this protein in the lifecycle of phages infecting Ralstonia solanacearum.

Main Methods:

  • In vivo recombination assays using E. coli and R. solanacearum plasmids.
  • Analysis of attP, attB, attL, and attR sequences.
  • Functional characterization of the ϕRSM1 integrase (ORF14).

Main Results:

  • The ϕRSM1 integrase (ORF14) mediates site-specific integration into the R. solanacearum attB sequence.
  • Intermolecular and intramolecular recombination events were observed in vivo.
  • The recombination products (attL and attR) matched those found in native prophages.

Conclusions:

  • The small serine recombinase ORF14 from ϕRSM1 functions in both integrative and excisive recombination.
  • This represents the first evidence of resolvase/invertase subfamily members mediating these processes in filamentous phages.

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