New reagent for discrimination of single- and double-stranded regions in DNA.
It was found that DNA alkylation at the N-7 guanine with the bulky alkylating reagent, N,N,N'-tri-(beta-chloroethyl)-N'-(p-formylphenyl)propylene diamine-1,3 (TFP) is much diminished when DNA is double-stranded. We report here an application of this reaction for probing the hairpin structure in the palindrome-containing single-stranded (ss) DNA fragment of 377 bases prepared from the Eco-RI-BaMHI fragment of plasmid pBR322. 5'-Labeled ss fragment was modified with TFP and cleaved by piperidine hydrolysis at the alkylated guanine residues according to the Maxam-Gilbert procedure. Guanines in the hairpin formed by palindrome of 9 bp were protected from TFP action, while dimethyl sulfate modified all guanines.
Mazin
A V
AV
Institute of Cytology and Genetics, Academy of Sciences of USSR, Novosibirsk.
Kuzminov
A V
AV
Dianov
G L
GL
Salganik
R I
RI
D000477:Alkylating Agents; D001483:Base Sequence; D004247:DNA; D004269:DNA, Bacterial; D004277:DNA, Single-Stranded; D004279:DNA, Viral; D008969:Molecular Sequence Data; D009588:Nitrogen Mustard Compounds; D009690:Nucleic Acid Conformation; D009691:Nucleic Acid Denaturation; D010957:Plasmids
D000477:Alkylating Agents; D004269:DNA, Bacterial; D004277:DNA, Single-Stranded; D004279:DNA, Viral; D009588:Nitrogen Mustard Compounds; C020534:N,N,N'-tri(beta-chloroethyl)-N'-(4-formylphenyl)-1,3-propylenediamine; D004247:DNA