Title:
METHOD OF SITE SPECIFICALLY INTRODUCING TYROSINE ANALOGUE INTO PROTEIN
Document Type and Number:
WIPO Patent Application WO/2006/135096
Kind Code:
A1
Abstract:
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 tRNATyr and the mutant tyrosyl-tRNA synthetase.
Inventors:
NISHIKAWA KAZUYA (JP)
YOKOGAWA TAKASHI (JP)
OHNO SATOSHI (JP)
YOKOGAWA TAKASHI (JP)
OHNO SATOSHI (JP)
Application Number:
PCT/JP2006/312370
Publication Date:
December 21, 2006
Filing Date:
June 14, 2006
Export Citation:
Assignee:
UNIV GIFU (JP)
NISHIKAWA KAZUYA (JP)
YOKOGAWA TAKASHI (JP)
OHNO SATOSHI (JP)
NISHIKAWA KAZUYA (JP)
YOKOGAWA TAKASHI (JP)
OHNO SATOSHI (JP)
International Classes:
C12P21/02; C12N15/09
Foreign References:
| JP2004261160A | 2004-09-24 | |||
| JPH05236987A | 1993-09-17 |
Other References:
MA C. ET AL.: "In vitro protein engineering using synthetic tRNA(Ala) with different anticodons", BIOCHEMISTRY, vol. 32, 1993, pages 7939 - 7945, XP003002661
CAYAMA E. ET AL.: "New chromatographic and biochemical strategies for quick preparative isolation of tRNA", NUCLEIC ACIDS RES., vol. 28, no. E64, 2000, XP003002662
KAO C. ET AL.: "A simple and efficient method to reduce nontemplated nucleotide addition at the 3 terminus of RNAs transcribed by T7 RNA polymerase", RNA, vol. 5, 1999, pages 1268 - 1272, XP003002663
WANG L. ET AL.: "Expanding the genetic code of Escherichia coli", SCIENCE, vol. 292, 20 April 2001 (2001-04-20), pages 498 - 500, XP002904367
CAYAMA E. ET AL.: "New chromatographic and biochemical strategies for quick preparative isolation of tRNA", NUCLEIC ACIDS RES., vol. 28, no. E64, 2000, XP003002662
KAO C. ET AL.: "A simple and efficient method to reduce nontemplated nucleotide addition at the 3 terminus of RNAs transcribed by T7 RNA polymerase", RNA, vol. 5, 1999, pages 1268 - 1272, XP003002663
WANG L. ET AL.: "Expanding the genetic code of Escherichia coli", SCIENCE, vol. 292, 20 April 2001 (2001-04-20), pages 498 - 500, XP002904367
Attorney, Agent or Firm:
Hiraki, Yusuke (3-20 Toranomon 4-chom, Minato-ku Tokyo 01, JP)
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