New reagent for discrimination of single- and double-stranded regions in DNA

A. V. Mazin; A. V. Kuzminov; G. L. Dianov; R. I. Salganik

doi:10.1016/0014-5793(89)81664-3

New reagent for discrimination of single- and double-stranded regions in DNA

A. V. Mazin, A. V. Kuzminov, G. L. Dianov, R. I. Salganik

Research output: Contribution to journal › Article › peer-review

Abstract

It was found that DNA alkylation at the N-7 guanine with the bulky alkylating reagent, N,N,N'-tri-(β-chloroethyl)-N-(p-formylpheny)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 EcoRI-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.

Original language	English (US)
Pages (from-to)	244-246
Number of pages	3
Journal	FEBS Letters
Volume	258
Issue number	2
DOIs	https://doi.org/10.1016/0014-5793(89)81664-3
State	Published - Dec 4 1989
Externally published	Yes

Keywords

Bulky alkylating reagent
Guanine alkylation, N-7
Structure probing, hairpin

ASJC Scopus subject areas

Biophysics
Structural Biology
Biochemistry
Molecular Biology
Genetics
Cell Biology

Online availability

10.1016/0014-5793(89)81664-3

Library availability

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title = "New reagent for discrimination of single- and double-stranded regions in DNA",

abstract = "It was found that DNA alkylation at the N-7 guanine with the bulky alkylating reagent, N,N,N'-tri-(β-chloroethyl)-N-(p-formylpheny)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 EcoRI-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.",

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doi = "10.1016/0014-5793(89)81664-3",

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T1 - New reagent for discrimination of single- and double-stranded regions in DNA

AU - Mazin, A. V.

AU - Kuzminov, A. V.

AU - Dianov, G. L.

AU - Salganik, R. I.

PY - 1989/12/4

Y1 - 1989/12/4

N2 - It was found that DNA alkylation at the N-7 guanine with the bulky alkylating reagent, N,N,N'-tri-(β-chloroethyl)-N-(p-formylpheny)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 EcoRI-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.

AB - It was found that DNA alkylation at the N-7 guanine with the bulky alkylating reagent, N,N,N'-tri-(β-chloroethyl)-N-(p-formylpheny)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 EcoRI-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.

KW - Bulky alkylating reagent

KW - Guanine alkylation, N-7

KW - Structure probing, hairpin

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New reagent for discrimination of single- and double-stranded regions in DNA

Abstract

Keywords

ASJC Scopus subject areas

Online availability

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