THE SCRIPPS RESEARCH INSTITUTE;THE REGENTS OF THE UNIVERSITY OF CALIFORNIA
发明人:
SCHULTZ, PETER,WANG, LEI,ANDERSON, JOHN CHRISTOPHER,CHIN, JASON W.,LIU, DAVID R.,MAGLIERY, THOMAS J.,MEGGERS, ERIC L.,MEHL, RYAN AARON,PASTRNAK, MIRO,SANTORO, STEPHEN WILLIAM,ZHANG, ZHIWEN
申请号：
ES09006800
公开号：
ES2464532T3
申请日:
2002.04.19
申请国别(地区):
ES
年份:
2014
代理人:
摘要：
A pair of aminoacyl tRNA synthetase and tRNA, which directs the incorporation of an unnatural amino acid into the amino acid sequence of a polypeptide, where the aminoacyl tRNA synthetase exhibits selectivity for the unnatural amino acid compared to any of the twenty common amino acids, said TRNA is specific for an amber stop codon in the mRNA encoding the polypeptide and the aminoacyl tRNA synthetase aminoacylates the tRNA with the unnatural amino acid, wherein said unnatural amino acid is selected from the group consisting of: for acetyl phenylalanine, for amino phenylalanine and for nitro phenylalanine, and where the aminoacyl tRNA synthetase can be obtained by a method comprising: creating a library of mutated aminoacyl tRNA synthetases of Methanococcus jannaschii； positively selecting from the library of aminoacyl tRNA synthetases mutated from Methanococcus jannaschii members that aminoacilen said tRNA in the presence of said unnatural amino acid and a natural amino acid, thereby providing a reserve of active mutated aminoacyl tRNA synthetases of Methanococcus jannaschii； select negatively from the aminoacil pool of active mutated tRNA synthetases of Methanococcus jannaschii aminoacyl tRNA syntheses mutants of Methanococcus jannaschii that aminoacilen said tRNA in the absence of the unnatural amino acid； and thereby obtaining the aminoacyl tRNA synthetase which preferably aminoacylates said tRNA with said unnatural amino acid.Un par de aminoacil ARNt sintetasa y ARNt, que dirige la incorporación de un aminoácido no natural en la secuencia de aminoácidos de un polipéptido, donde la aminoacil ARNt sintetasa presenta selectividad por el aminoácido no natural en comparación con cualquiera de los veinte aminoácidos comunes, dicho ARNt es específico para un codón de parada ámbar en el ARNm que codifica el polipéptido y la aminoacil ARNt sintetasa aminoacila el ARNt con el aminoácido no natural, donde dicho aminoácido no natural se selecciona entre el grupo que consis
