Recombination of synthetic oligonucleotides with prokaryotic chromosomes: Substrate requirements of the Escherichia coli/λrd and Sulfolobus acidocaldarius recombination systems

Dennis W. Grogan, Kristy R. Stengel

Research output: Contribution to journalArticlepeer-review

34 Scopus citations

Abstract

In order to reveal functional properties of recombination involving short ssDNAs in hyperthermophilic archaea, we evaluated oligonucleotide-mediated transformation (OMT) in Sulfolobus acidocaldarius and Escherichia coli as a function of the molecular properties of the ssDNA substrates. Unmodified ssDNAs as short as 20-22 nt yielded recombinants in both organisms, as did longer DNAs forming as few as 2-5 base pairs on one side of the genomic mutation. The two OMT systems showed similar responses to certain end modifications of the oligonucleotides, but E. coli was found to require a 5′ phosphate on 5′-limited ssDNA whereas this requirement was not evident in S. acidocaldarius. The ability of both E. coli and S. acidocaldarius to incorporate short, mismatched ssDNAs into their genomes raises questions about the biological significance of this capability, including its phylogenetic distribution among microorganisms and its impact on genome stability. These questions seem particularly relevant for S. acidocaldarius, as this archaeon has natural competence for OMT, encodes no MutSL homologues and thrives under environmental conditions that accelerate DNA decomposition.

Original languageEnglish (US)
Pages (from-to)1255-1265
Number of pages11
JournalMolecular Microbiology
Volume69
Issue number5
DOIs
StatePublished - Sep 2008
Externally publishedYes

ASJC Scopus subject areas

  • Microbiology
  • Molecular Biology

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