Use of polymerase chain reaction catalyzed by Taq DNA polymerase for site-specific mutagenesis.
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.
Kadowaki
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Diabetes Branch, National Institute of Diabetes and Digestive and Kidney Diseases, Bethesda, MD.
Kadowaki
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Wondisford
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Taylor
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D001483:Base Sequence; D002384:Catalysis; D055598:Chemical Phenomena; D002621:Chemistry; D003001:Cloning, Molecular; D004274:DNA, Recombinant; D004259:DNA-Directed DNA Polymerase / Q000378:metabolism*; D005784:Gene Amplification; D005822:Genetic Vectors; D009154:Mutation; D009838:Oligodeoxyribonucleotides / Q000096:biosynthesis / Q000235:genetics; D010957:Plasmids; D011972:Receptor, Insulin / Q000235:genetics; D019914:Taq Polymerase; D013698:Templates, Genetic
D004274:DNA, Recombinant; D009838:Oligodeoxyribonucleotides; D011972:Receptor, Insulin; D019914:Taq Polymerase; D004259:DNA-Directed DNA Polymerase