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EMBO J
1984 Nov 01;311:2445-52. doi: 10.1002/j.1460-2075.1984.tb02154.x.
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Synthesis of an ochre suppressor tRNA gene and expression in mammalian cells.
Laski FA
,
Belagaje R
,
Hudziak RM
,
Capecchi MR
,
Norton GP
,
Palese P
,
RajBhandary UL
,
Sharp PA
.
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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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