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New methods enabling efficient incorporation of unnatural amino acids in yeast.

  • Academic Journal
  • Wang Q; Salk Institute for Biological Studies, 10010 North Torrey Pines Road, La Jolla, California 92037, USA.
    Wang L
  • Journal of the American Chemical Society [J Am Chem Soc] 2008 May 14; Vol. 130 (19), pp. 6066-7. Date of Electronic Publication: 2008 Apr 22.
  • English
  • New chemical and physical properties can be selectively introduced into proteins directly in live cells by genetically incorporating unnatural amino acids. The incorporation efficiency determines how effective such properties can be exploited and was very low in yeast. We developed a new method for efficient expression of orthogonal bacterial tRNA in yeast using polymerase III promoters that are cleaved from primary transcripts. In addition, a yeast strain deficient in nonsense-mediated mRNA decay was generated to prevent rapid degradation of target mRNA containing premature stop codons, which are the most frequently used to encode unnatural amino acids. These new strategies enabled a significant increase in yield of unnatural amino acid containing proteins from tens of micrograms to tens of milligrams per liter in yeast.
Additional Information
Publisher: American Chemical Society Country of Publication: United States NLM ID: 7503056 Publication Model: Print-Electronic Cited Medium: Internet ISSN: 1520-5126 (Electronic) Linking ISSN: 00027863 NLM ISO Abbreviation: J Am Chem Soc Subsets: MEDLINE
Publication: Washington, DC : American Chemical Society
Original Publication: Easton, Pa. [etc.]
0 (Amino Acids)
0 (RNA, Messenger)
9014-25-9 (RNA, Transfer)
Date Created: 20080423 Date Completed: 20080530 Latest Revision: 20081216

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