A novel method for converting common restriction enzymes into rare cutters: integration host factor-mediated Achilles' cleavage (IHF-AC)

Jósef Kur, Michael Koob, Adam Burkiewicz, Waclaw Szybalski

Research output: Contribution to journalArticlepeer-review

8 Scopus citations

Abstract

Integration host factor (IHF)-mediated protection against enzymatic methylation at ihf-overlapping sites provides the basis for this novel application of the Achilles' cleavage (AC) technique [Koob., Science 241 (1998) 1084-10861 for generating rare natural cleavage sites. When applying IHF-AC to plasmid, phage λ, Escherichia coli and yeast genomes, only a few of the EcoRI, MboI sites (sites overlapped the ihf sites)remained cleavable after prior methylation with the cognate M· EcoRI, M· HinfI, or Dam methyltransferases in the presence of IHF. Thus, IHF-AC essentially converted these enzymes into very rare cutters. The extent of cleavage could be controlled by varying the IHF:DNA ratio and temperature. Moreover, the method permits the genomic location and strength of the ihf sites to be determined.

Original languageEnglish (US)
Pages (from-to)1-7
Number of pages7
JournalGene
Volume110
Issue number1
DOIs
StatePublished - Jan 2 1992

Keywords

  • (Methyltransferases
  • DNA-binding protein
  • Dam
  • EcoRI
  • HinfO
  • pulsed-field gel electrophoresis
  • recombinant DNA)

Fingerprint Dive into the research topics of 'A novel method for converting common restriction enzymes into rare cutters: integration host factor-mediated Achilles' cleavage (IHF-AC)'. Together they form a unique fingerprint.

Cite this