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Expression of a Set of Synthetic Suppressor tRNAPhe Genes in Saccharomyces cerevisiae

Masson, Jean-Michel and Meuris, Pierre and Grunstein, Michael and Abelson, John and Miller, Jeffrey H. (1987) Expression of a Set of Synthetic Suppressor tRNAPhe Genes in Saccharomyces cerevisiae. Proceedings of the National Academy of Sciences of the United States of America, 84 (19). pp. 6815-6819. ISSN 0027-8424. PMCID PMC299175. doi:10.1073/pnas.84.19.6815.

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ynthetic ochre and amber tRNA suppressor genes derived from the yeast tRNAGAAPhe sequence have been constructed. They were efficiently transcribed in vitro and expressed in vivo via a synthetic expression cassette. tRNAUUAPhe and tRNAUUAPheDelta IVS (IVS = intervening sequence) are relatively inefficient ochre suppressors. They are toxic to the cell when expressed on a multicopy plasmid, and they do not suppress at all when present as single copies. The intron does not seem to have any effect on suppression. In contrast, the amber suppressor tRNACUAPheDelta IVS is efficient when expressed from a single-copy plasmid, while its efficiency is reduced on a multicopy vector.

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Additional Information:© 1987 by the National Academy of Sciences Contributed by John Abelson, June 15, 1987 We thank Susan Liebman and Maynard Olson for providing strains SL183-21c and AHA75, respectively; Sandra Ebeling for technical assistance; Marge Strobel for help with the in vitro reactions; and Susan Whoriskey for the DNA sequencing. This work was supported in part by a National Science Foundation Grant (DMB 8417353) to J.A. and J.H.M. and by a National Institutes of Health Grant (GM23674) to M.G. The authors are indebted to the Biotechnology Research and Education Program for use of equipment. The publication costs of this article were defrayed in part by page charge payment. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. §1734 solely to indicate this fact.
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Issue or Number:19
PubMed Central ID:PMC299175
Record Number:CaltechAUTHORS:MASpnas87
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ID Code:6367
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Deposited On:05 Dec 2006
Last Modified:08 Nov 2021 20:33

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