METHOD OF SITE SPECIFICALLY INTRODUCING TYROSINE ANALOGUE INTO PROTEIN
摘要
<p>It is intended to provide a method of producing a mutated protein containing a tyrosine analogue in codons coding for amino acids other than stop codons such as an amber codon without introducing any mutation into a target gene. (i) A mutation is introduced into tyrosine tRNA derived from yeast to modify it for AGU;(ii) a mutant tyrosyl-tRNA synthetase derived from yeast is utilized; (iii) all types of tRNA were selectively isolated form an E. coli cell extract protein synthesis system, only tRNA for AGU is removed from the all types of tRNA and the tRNA other than the tRNA for AGU are returned to an E. coli-derived cell-free protein synthesis system. It was found that an unnatural amino acid-containing protein can be synthesized by the E. coli-derived cell-free protein synthesis system from which the tRNA for AGU has been removed using the modified tRNA<SUP>Tyr</SUP> and the mutant tyrosyl-tRNA synthetase.</p>
