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Synthesis of an ochre suppressor tRNA gene and expression in mammalian cells.

Authors
  • Laski, F A
  • Belagaje, R
  • Hudziak, R M
  • Capecchi, M R
  • Norton, G P
  • Palese, P
  • RajBhandary, U L
  • Sharp, P A
Type
Published Article
Journal
The EMBO journal
Publication Date
Nov 01, 1984
Volume
3
Issue
11
Pages
2445–2452
Identifiers
PMID: 6096120
Source
Medline
License
Unknown

Abstract

We have used site-specific mutagenesis to change the anticodon of a Xenopus laevis tyrosine tRNA gene so that it would recognize ochre codons. This tRNA gene is expressed when amplified in monkey cells as part of a SV40 recombinant and efficiently suppresses termination at both the ochre codon separating the adenovirus 2 hexon gene from a 23-kd downstream gene and the ochre codon at the end of the NS1 gene of influenza virus A/Tex/1/68. Termination at an amber codon of a NS1 gene of another influenza virus strain was not suppressed by the (Su+) ochre gene suggesting that in mammalian cells amber codons are not recognized by ochre suppressor tRNAs. Finally, microinjection into mammalian cells of both (Su+) ochre tRNA genes and selectible genes containing ochre nonsense mutations gives rise to colonies under selective conditions. We conclude that it should be possible to isolate a wide assortment of mammalian cell lines with ochre suppressor activity.

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