Synthesis of an Ochre Suppressor TRNA Gene and Expression in Mammalian Cells

Overview

Journal EMBO J

Specialties Biotechnology
Molecular Biology

Date 1984 Nov 1

PMID 6096120

Citations 15

Authors

F A Laski

R Belagaje

R M Hudziak

M R Capecchi

G P Norton

P Palese

U L RajBhandary

P A Sharp

Affiliations

Soon will be listed here.

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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