Multisite oligonucleotide-mediated mutagenesis: application to the conversion of a mitochondrial gene to universal genetic code.
Using multisite oligonucleotide-mediated mutagenesis in conjunction with a mutagenesis selection procedure and rapid screening by allele-specific oligonucleotide hybridization has allowed us to develop a reliable protocol that enables a large number of base changes to be introduced rapidly into a piece of DNA, with the minimum number of manipulations. We have applied this protocol to generate synthetic versions of four mouse mitochondrial genes capable of being expressed in the nucleus/cytosol.
Sutherland
L
L
University of Glasgow, Scotland, UK.
Davidson
J
J
Glass
L L
LL
Jacobs
H T
HT
D000595:Amino Acid Sequence; D000818:Animals; D017104:Bacteriophage M13 / Q000235:genetics; D001483:Base Sequence; D003001:Cloning, Molecular; D004272:DNA, Mitochondrial / Q000235:genetics*; D003576:Electron Transport Complex IV / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics; D005813:Genes, Synthetic; D005815:Genetic Code; D051379:Mice; D008969:Molecular Sequence Data; D016296:Mutagenesis; D006180:Proton-Translocating ATPases / Q000235:genetics
D004272:DNA, Mitochondrial; D003576:Electron Transport Complex IV; D006180:Proton-Translocating ATPases