TY - JOUR
T1 - A novel method for converting common restriction enzymes into rare cutters
T2 - integration host factor-mediated Achilles' cleavage (IHF-AC)
AU - Kur, Jósef
AU - Koob, Michael
AU - Burkiewicz, Adam
AU - Szybalski, Waclaw
PY - 1992/1/2
Y1 - 1992/1/2
N2 - 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.
AB - 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.
KW - (Methyltransferases
KW - DNA-binding protein
KW - Dam
KW - EcoRI
KW - HinfO
KW - pulsed-field gel electrophoresis
KW - recombinant DNA)
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U2 - 10.1016/0378-1119(92)90437-T
DO - 10.1016/0378-1119(92)90437-T
M3 - Article
C2 - 1531969
AN - SCOPUS:0026565305
SN - 0378-1119
VL - 110
SP - 1
EP - 7
JO - Gene
JF - Gene
IS - 1
ER -