Use of polymerase chain reaction catalyzed by Taq DNA polymerase for site-specific mutagenesis

Hiroko Kadowaki; Takashi Kadowaki; Fredric E. Wondisford; Simeon I. Taylor

doi:10.1016/0378-1119(89)90018-8

Use of polymerase chain reaction catalyzed by Taq DNA polymerase for site-specific mutagenesis

Hiroko Kadowaki, Takashi Kadowaki, Fredric E. Wondisford, Simeon I. Taylor

School of Medicine

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

94 Scopus citations

Abstract

The polymerase chain reaction catalyzed by Taq DNA polymerase has been used for site-specific mutagenesis. The amplification was primed by two oligodeoxyribonucleotides complementary to insulin receptor cDNA. To direct the synthesis of mutant DNA, mismatches were introduced into one of the primers. Six different mutations were constructed by this technique. Of twelve clones whose sequences were determined, ten (83%) had the correct sequence. This technique, which does not require the use of single-stranded DNA templates, provides a simple and efficient approach to site-specific mutagenesis.

Original language	English (US)
Pages (from-to)	161-166
Number of pages	6
Journal	Gene
Volume	76
Issue number	1
DOIs	https://doi.org/10.1016/0378-1119(89)90018-8
State	Published - Mar 15 1989

Keywords

Recombinant DNA
cloning
gene amplification
insulin receptor
oligodeoxyribonucleotide primers
plasmid

ASJC Scopus subject areas

Genetics

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10.1016/0378-1119(89)90018-8

Cite this

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title = "Use of polymerase chain reaction catalyzed by Taq DNA polymerase for site-specific mutagenesis",

abstract = "The polymerase chain reaction catalyzed by Taq DNA polymerase has been used for site-specific mutagenesis. The amplification was primed by two oligodeoxyribonucleotides complementary to insulin receptor cDNA. To direct the synthesis of mutant DNA, mismatches were introduced into one of the primers. Six different mutations were constructed by this technique. Of twelve clones whose sequences were determined, ten (83%) had the correct sequence. This technique, which does not require the use of single-stranded DNA templates, provides a simple and efficient approach to site-specific mutagenesis.",

keywords = "Recombinant DNA, cloning, gene amplification, insulin receptor, oligodeoxyribonucleotide primers, plasmid",

author = "Hiroko Kadowaki and Takashi Kadowaki and Wondisford, {Fredric E.} and Taylor, {Simeon I.}",

note = "Funding Information: Dr. Hiroko Kadowaki is supported by the American Diabetes Association through their Mentor-Based Fellowship Program. Partial research support was also generously provided by a research grant from Juvenile Diabetes Foundation International.",

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AU - Kadowaki, Hiroko

AU - Kadowaki, Takashi

AU - Wondisford, Fredric E.

AU - Taylor, Simeon I.

N1 - Funding Information: Dr. Hiroko Kadowaki is supported by the American Diabetes Association through their Mentor-Based Fellowship Program. Partial research support was also generously provided by a research grant from Juvenile Diabetes Foundation International.

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Y1 - 1989/3/15

N2 - The polymerase chain reaction catalyzed by Taq DNA polymerase has been used for site-specific mutagenesis. The amplification was primed by two oligodeoxyribonucleotides complementary to insulin receptor cDNA. To direct the synthesis of mutant DNA, mismatches were introduced into one of the primers. Six different mutations were constructed by this technique. Of twelve clones whose sequences were determined, ten (83%) had the correct sequence. This technique, which does not require the use of single-stranded DNA templates, provides a simple and efficient approach to site-specific mutagenesis.

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KW - Recombinant DNA

KW - cloning

KW - gene amplification

KW - insulin receptor

KW - oligodeoxyribonucleotide primers

KW - plasmid

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Use of polymerase chain reaction catalyzed by Taq DNA polymerase for site-specific mutagenesis

Abstract

Keywords

ASJC Scopus subject areas

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