Simple and efficient oligonucleotide-directed mutagenesis using one primer and circular plasmid DNA template.
A rapid and simple procedure for site-directed mutagenesis is described. This method uses only a single oligonucleotide primer with the double-stranded circular plasmid DNA as the template for mutagenesis. The phage T4 gene 32 product is included during primer extension in vitro to increase efficiency. Single and multiple changes as well as deletions have been obtained at an efficiency of 1-2%.
Marotti
K R
KR
Molecular Biology Research Department, Upjohn Company, Kalamazoo, Michigan 49007.
Tomich
C S
CS
D000818:Animals; D001483:Base Sequence; D002417:Cattle; D004270:DNA, Circular / Q000235:genetics*; D004268:DNA-Binding Proteins / Q000235:genetics; D004352:Drug Resistance, Microbial / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics; D005821:Genetic Techniques; D013006:Growth Hormone / Q000235:genetics; D007612:Kanamycin / Q000494:pharmacology; D008969:Molecular Sequence Data; D009154:Mutation; D009841:Oligonucleotides / Q000235:genetics*; D010957:Plasmids; D013604:T-Phages / Q000235:genetics; D013698:Templates, Genetic; D010959:Tissue Plasminogen Activator / Q000235:genetics; D014170:Transformation, Genetic; D014764:Viral Proteins / Q000235:genetics
D004270:DNA, Circular; D004268:DNA-Binding Proteins; D009841:Oligonucleotides; D014764:Viral Proteins; C060035:gp32 protein, Enterobacteria phage T4; D007612:Kanamycin; D013006:Growth Hormone; D010959:Tissue Plasminogen Activator