Nonnatural mutagenesis in E. coli and rabbit reticulocyte lysates by using four-base codons.
Various four-base codons had been developed for introduction of nonnatural amino acids into proteins in an E. coli in vitro translation system. Here, the four-base codon mediated incorporation of nonnatural amino acids was investigated in a rabbit reticulocyte lysate. Mutated streptavidin mRNAs containing four-base codons were prepared and expressed in a rabbit reticulocyte lysate in the presence of tRNAs containing corresponding four-base anticodons. Western blot analysis indicates that various four-base codons are efficiently decoded by the corresponding tRNAs in the rabbit in vitro translation system.
Hohsaka
Takahiro
T
Department of Bioscience and Biotechnology, Okayama University, Okayama 700-8530, Japan.
Fukushima
Masaharu
M
Sisido
Masahiko
M
D000595:Amino Acid Sequence; D000818:Animals; D003062:Codon; D004926:Escherichia coli / Q000235:genetics*; D016296:Mutagenesis; D011817:Rabbits
D003062:Codon