Title:
METHOD OF EXPRESSING PROTEIN HAVING UNNATURAL AMINO ACID INTEGRATED THEREINTO
Document Type and Number:
WIPO Patent Application WO/2004/039989
Kind Code:
A1
Abstract:
A method of expressing a protein having an unnatural amino acid integrated thereinto characterized by comprising expressing (A) a mutant tyrosyl tRNA synthase which is a mutant of Escherichia coli-origin tyrosyl tRNA synthase and has an elevated specificity for a unnatural tyrosine derivative compared with the specificity for tyrosine, (B) a suppressor tRNA originating in an eubacterium belonging to the genus Bacillus, Mycoplasma or Staphylococcus which is capable of binding to the above tyrosine derivative in the presence of the above mutant TyrRS, and (C) a desired protein gene having a nonsense mutation at a desired site in animal cells to thereby incorporate the above tyrosine derivative into the nonsense mutation site in the above protein.
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Inventors:
YOKOYAMA SHIGEYUKI (JP)
SHIROUZU MIKAKO (JP)
SAKAMOTO AYAKO (JP)
SAKAMOTO KENSAKU (JP)
SHIROUZU MIKAKO (JP)
SAKAMOTO AYAKO (JP)
SAKAMOTO KENSAKU (JP)
Application Number:
PCT/JP2003/014028
Publication Date:
May 13, 2004
Filing Date:
October 31, 2003
Export Citation:
Assignee:
RIKEN (JP)
JAPAN SCIENCE & TECH AGENCY (JP)
YOKOYAMA SHIGEYUKI (JP)
SHIROUZU MIKAKO (JP)
SAKAMOTO AYAKO (JP)
SAKAMOTO KENSAKU (JP)
JAPAN SCIENCE & TECH AGENCY (JP)
YOKOYAMA SHIGEYUKI (JP)
SHIROUZU MIKAKO (JP)
SAKAMOTO AYAKO (JP)
SAKAMOTO KENSAKU (JP)
International Classes:
C12N9/00; A61K48/00; (IPC1-7): C12N15/85; C12N5/10
Other References:
SAKAMOTO K ET AL: "Site-specific incorporation of an unnatural amino acid into proteins in mammalian cells", NUCLEIC ACIDS RES, vol. 30, no. 21, 1 November 2002 (2002-11-01), pages 4692 - 4699, XP002976283
DAISUKE KIGA ET AL: "Hen'igata daichokin Tyrosyl-tRNA Gosei Koso o Riyo Shita, Shinkaku Seibutsu no Musaibo Honyakukei ni okeru 3-Iodotyrosine no Tampaku-shitsu eno Bui Tokuiteki torikumi", SEI KAGAKU, vol. 74, no. 8, 25 August 2002 (2002-08-25), pages 1011, XP002976284
KIGA D ET AL: "An engineered Escherichia coli tyrosyl-tRNA synthetase for site-specific incorporation of an unnatural amino acid into proteins in eukaryotic translation and its application in a wheat germ cell-free system", PROC.NATL.ACAD.SCI.USA, vol. 99, no. 15, 23 July 2002 (2002-07-23), pages 9715 - 9720, XP002976285
WAWROUSEK EF ET AL.: "Two large clusters with thirty-seven transfer RNA genes adjacent to ribosomal RNA gene sets in Bacillus subtilis. Sequence and organization of trrnD and trrnE gene clusters", J.BIOL.CHEM, vol. 259, no. 6, 1984, pages 3694 - 3702, XP002976286
DAISUKE KIGA ET AL: "Hen'igata daichokin Tyrosyl-tRNA Gosei Koso o Riyo Shita, Shinkaku Seibutsu no Musaibo Honyakukei ni okeru 3-Iodotyrosine no Tampaku-shitsu eno Bui Tokuiteki torikumi", SEI KAGAKU, vol. 74, no. 8, 25 August 2002 (2002-08-25), pages 1011, XP002976284
KIGA D ET AL: "An engineered Escherichia coli tyrosyl-tRNA synthetase for site-specific incorporation of an unnatural amino acid into proteins in eukaryotic translation and its application in a wheat germ cell-free system", PROC.NATL.ACAD.SCI.USA, vol. 99, no. 15, 23 July 2002 (2002-07-23), pages 9715 - 9720, XP002976285
WAWROUSEK EF ET AL.: "Two large clusters with thirty-seven transfer RNA genes adjacent to ribosomal RNA gene sets in Bacillus subtilis. Sequence and organization of trrnD and trrnE gene clusters", J.BIOL.CHEM, vol. 259, no. 6, 1984, pages 3694 - 3702, XP002976286
Attorney, Agent or Firm:
Shiga, Masatake (Yaesu Chuo-ku, Tokyo, JP)
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