In vitro mutagenesis by incorporation of N4-aminodeoxycytidine 5'-triphosphate.

K. Negishi, K. Matsumoto, T. Bessho, F. Tada, H. Hayatsu

Research output: Contribution to journalArticle

3 Citations (Scopus)

Abstract

We have investigated the possibility of introducing a new way to carry out in vitro mutagenesis. N4-Aminodeoxycytidine 5'-triphosphate was used in the Klenow enzyme-catalyzed chain elongation of a primer oligonucleotide hybridized to a lacZ alpha region of M13mp2 viral single strand DNA, and the possibility of inducing an efficient, randomly distributed point mutations into this particular genomic region was explored. On transfection of the resulting DNA into E. coli, mutant phages emerged at frequencies up to 1%. Analysis of the DNA sequences of the mutants has shown that single transitions, either A to G or G to A, were induced in a random fashion, thus providing data to show the possibility of using this method for production of mutant proteins having various single amino-acid changes in a defined domain.

Original languageEnglish (US)
Pages (from-to)33-36
Number of pages4
JournalNucleic acids symposium series
Issue number19
StatePublished - 1988

Fingerprint

Mutagenesis
DNA Primers
DNA
Mutant Proteins
DNA Sequence Analysis
Point Mutation
Bacteriophages
Transfection
Escherichia coli
Amino Acids
Enzymes
In Vitro Techniques
triphosphoric acid

ASJC Scopus subject areas

  • Medicine(all)

Cite this

In vitro mutagenesis by incorporation of N4-aminodeoxycytidine 5'-triphosphate. / Negishi, K.; Matsumoto, K.; Bessho, T.; Tada, F.; Hayatsu, H.

In: Nucleic acids symposium series, No. 19, 1988, p. 33-36.

Research output: Contribution to journalArticle

Negishi, K. ; Matsumoto, K. ; Bessho, T. ; Tada, F. ; Hayatsu, H. / In vitro mutagenesis by incorporation of N4-aminodeoxycytidine 5'-triphosphate. In: Nucleic acids symposium series. 1988 ; No. 19. pp. 33-36.
@article{639c1884cb694d32abaa45935c0ff380,
title = "In vitro mutagenesis by incorporation of N4-aminodeoxycytidine 5'-triphosphate.",
abstract = "We have investigated the possibility of introducing a new way to carry out in vitro mutagenesis. N4-Aminodeoxycytidine 5'-triphosphate was used in the Klenow enzyme-catalyzed chain elongation of a primer oligonucleotide hybridized to a lacZ alpha region of M13mp2 viral single strand DNA, and the possibility of inducing an efficient, randomly distributed point mutations into this particular genomic region was explored. On transfection of the resulting DNA into E. coli, mutant phages emerged at frequencies up to 1{\%}. Analysis of the DNA sequences of the mutants has shown that single transitions, either A to G or G to A, were induced in a random fashion, thus providing data to show the possibility of using this method for production of mutant proteins having various single amino-acid changes in a defined domain.",
author = "K. Negishi and K. Matsumoto and T. Bessho and F. Tada and H. Hayatsu",
year = "1988",
language = "English (US)",
pages = "33--36",
journal = "Nucleic acids symposium series",
issn = "0261-3166",
publisher = "Information Retrieval",
number = "19",

}

TY - JOUR

T1 - In vitro mutagenesis by incorporation of N4-aminodeoxycytidine 5'-triphosphate.

AU - Negishi, K.

AU - Matsumoto, K.

AU - Bessho, T.

AU - Tada, F.

AU - Hayatsu, H.

PY - 1988

Y1 - 1988

N2 - We have investigated the possibility of introducing a new way to carry out in vitro mutagenesis. N4-Aminodeoxycytidine 5'-triphosphate was used in the Klenow enzyme-catalyzed chain elongation of a primer oligonucleotide hybridized to a lacZ alpha region of M13mp2 viral single strand DNA, and the possibility of inducing an efficient, randomly distributed point mutations into this particular genomic region was explored. On transfection of the resulting DNA into E. coli, mutant phages emerged at frequencies up to 1%. Analysis of the DNA sequences of the mutants has shown that single transitions, either A to G or G to A, were induced in a random fashion, thus providing data to show the possibility of using this method for production of mutant proteins having various single amino-acid changes in a defined domain.

AB - We have investigated the possibility of introducing a new way to carry out in vitro mutagenesis. N4-Aminodeoxycytidine 5'-triphosphate was used in the Klenow enzyme-catalyzed chain elongation of a primer oligonucleotide hybridized to a lacZ alpha region of M13mp2 viral single strand DNA, and the possibility of inducing an efficient, randomly distributed point mutations into this particular genomic region was explored. On transfection of the resulting DNA into E. coli, mutant phages emerged at frequencies up to 1%. Analysis of the DNA sequences of the mutants has shown that single transitions, either A to G or G to A, were induced in a random fashion, thus providing data to show the possibility of using this method for production of mutant proteins having various single amino-acid changes in a defined domain.

UR - http://www.scopus.com/inward/record.url?scp=0024194671&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0024194671&partnerID=8YFLogxK

M3 - Article

C2 - 3067218

AN - SCOPUS:0024194671

SP - 33

EP - 36

JO - Nucleic acids symposium series

JF - Nucleic acids symposium series

SN - 0261-3166

IS - 19

ER -