Details

Autor(en) / Beteiligte

• Laski, F.A.
• Belagaje, R.
• Hudziak, R.M.
• Capecchi, M.R.
• Norton, G.P.
• Palese, P.
• RajBhandary, U.L.
• Sharp, P.A.

Titel

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

Ist Teil von

• The EMBO journal, 1984-11, Vol.3 (11), p.2445-2452

Ort / Verlag

England

Erscheinungsjahr

1984

Quelle

Electronic Journals Library

Beschreibungen/Notizen

• 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.