Sequence specific inhibition of DNA restriction enzyme cleavage by PNA.
Plasmids containing double-stranded 10-mer PNA (peptide nucleic acid chimera) targets proximally flanked by two restriction enzyme sites were challenged with the complementary PNA or PNAs having one or two mismatches, and the effect on the restriction enzyme cleavage of the flanking sites was assayed. The following PNAs were used: T10-LysNH2, T5CT4-LysNH2 and T2CT2CT4-LysNH2 and the corresponding targets cloned into pUC 19 were flanked by BamH1, Sal1 or Pstl sites, respectively. In all cases it was found that complete inhibition of restriction enzyme cleavage was obtained with the complementary PNA, a significantly reduced effect was seen with a PNA having one mismatch, and no effect was seen with a PNA having two mismatches. These results show that PNA can be used as sequence specific blockers of DNA recognizing proteins.
Department of Biochemistry B, Panum Institute, Copenhagen, Denmark.
P E
PE
Nielsen
M
M
Egholm
R H
RH
Berg
O
O
Buchardt
D001483:Base Sequence; D002678:Chimera; D004247:DNA; D004262:DNA Restriction Enzymes / Q000037:antagonists & inhibitors*; D015245:Deoxyribonuclease BamHI / Q000037:antagonists & inhibitors; D015252:Deoxyribonucleases, Type II Site-Specific / Q000037:antagonists & inhibitors; D008969:Molecular Sequence Data; D009838:Oligodeoxyribonucleotides / Q000378:metabolism*; D010455:Peptides / Q000378:metabolism; D015183:Restriction Mapping
D009838:Oligodeoxyribonucleotides; D010455:Peptides; D004247:DNA; D004262:DNA Restriction Enzymes; D015245:Deoxyribonuclease BamHI; C024388:CTGCAG-specific type II deoxyribonucleases; D015252:Deoxyribonucleases, Type II Site-Specific; C029792:GTCGAC-specific type II deoxyribonucleases