Analysis of phage M13mp2 mutants produced from transfection of phage DNA having N4-aminocytosines at defined sequence positions

Keiko Matsumoto, Tetsuya Yashiki, Tadayoshi Bessho, Kazuo Negishi, Hikoya Hayatsu

Research output: Contribution to journalArticle

1 Citation (Scopus)

Abstract

N4-Aminocytidine is mutagenic in various organisms. In the cell, this cytidine analog is metabolized into N4-aminodeoxycytidine 5′-triphosphate, which will then be incorporated into DNA and mutation will result during the replication of the DNA. To prove that the N4-aminocytosine residue in DNA is indeed the site of mutagenesis, we prepared a series of phage M13mp2 DNA samples that bear N4-aminocytosine residues at a few defined positions in the lacZα region, by carrying out in vitro limited extension of primed phage DNA. Wh then transfected the DNAs to Escherichia coli and examined the progeny phages for the forward mutations. The M13mp2 DNAs bearing N4-aminocytosines produced mutant phages at high frequencies. Furthermore, DNA sequencing of the resulting mutants demonstrated that both AT-to-GC and GC-to-AT mutation took place at those position where N4-aminocytosine residues were originally present.

Original languageEnglish (US)
Pages (from-to)59-64
Number of pages6
JournalMutation Research - Fundamental and Molecular Mechanisms of Mutagenesis
Volume268
Issue number1
DOIs
StatePublished - Jul 1992

Fingerprint

Bacteriophages
Transfection
DNA
Mutation
Cytidine
DNA Replication
DNA Sequence Analysis
Mutagenesis
Escherichia coli

Keywords

  • DNA polymerase I
  • N-Aminodeoxycytidine 5′-triphosphate
  • Nucleoside analog
  • Site-directed mutagenesis

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics
  • Health, Toxicology and Mutagenesis

Cite this

Analysis of phage M13mp2 mutants produced from transfection of phage DNA having N4-aminocytosines at defined sequence positions. / Matsumoto, Keiko; Yashiki, Tetsuya; Bessho, Tadayoshi; Negishi, Kazuo; Hayatsu, Hikoya.

In: Mutation Research - Fundamental and Molecular Mechanisms of Mutagenesis, Vol. 268, No. 1, 07.1992, p. 59-64.

Research output: Contribution to journalArticle

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abstract = "N4-Aminocytidine is mutagenic in various organisms. In the cell, this cytidine analog is metabolized into N4-aminodeoxycytidine 5′-triphosphate, which will then be incorporated into DNA and mutation will result during the replication of the DNA. To prove that the N4-aminocytosine residue in DNA is indeed the site of mutagenesis, we prepared a series of phage M13mp2 DNA samples that bear N4-aminocytosine residues at a few defined positions in the lacZα region, by carrying out in vitro limited extension of primed phage DNA. Wh then transfected the DNAs to Escherichia coli and examined the progeny phages for the forward mutations. The M13mp2 DNAs bearing N4-aminocytosines produced mutant phages at high frequencies. Furthermore, DNA sequencing of the resulting mutants demonstrated that both AT-to-GC and GC-to-AT mutation took place at those position where N4-aminocytosine residues were originally present.",
keywords = "DNA polymerase I, N-Aminodeoxycytidine 5′-triphosphate, Nucleoside analog, Site-directed mutagenesis",
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AU - Yashiki, Tetsuya

AU - Bessho, Tadayoshi

AU - Negishi, Kazuo

AU - Hayatsu, Hikoya

PY - 1992/7

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N2 - N4-Aminocytidine is mutagenic in various organisms. In the cell, this cytidine analog is metabolized into N4-aminodeoxycytidine 5′-triphosphate, which will then be incorporated into DNA and mutation will result during the replication of the DNA. To prove that the N4-aminocytosine residue in DNA is indeed the site of mutagenesis, we prepared a series of phage M13mp2 DNA samples that bear N4-aminocytosine residues at a few defined positions in the lacZα region, by carrying out in vitro limited extension of primed phage DNA. Wh then transfected the DNAs to Escherichia coli and examined the progeny phages for the forward mutations. The M13mp2 DNAs bearing N4-aminocytosines produced mutant phages at high frequencies. Furthermore, DNA sequencing of the resulting mutants demonstrated that both AT-to-GC and GC-to-AT mutation took place at those position where N4-aminocytosine residues were originally present.

AB - N4-Aminocytidine is mutagenic in various organisms. In the cell, this cytidine analog is metabolized into N4-aminodeoxycytidine 5′-triphosphate, which will then be incorporated into DNA and mutation will result during the replication of the DNA. To prove that the N4-aminocytosine residue in DNA is indeed the site of mutagenesis, we prepared a series of phage M13mp2 DNA samples that bear N4-aminocytosine residues at a few defined positions in the lacZα region, by carrying out in vitro limited extension of primed phage DNA. Wh then transfected the DNAs to Escherichia coli and examined the progeny phages for the forward mutations. The M13mp2 DNAs bearing N4-aminocytosines produced mutant phages at high frequencies. Furthermore, DNA sequencing of the resulting mutants demonstrated that both AT-to-GC and GC-to-AT mutation took place at those position where N4-aminocytosine residues were originally present.

KW - DNA polymerase I

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KW - Nucleoside analog

KW - Site-directed mutagenesis

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