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PMID: 6096120 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S. Research Support, U.S. Gov't, P.H.S.

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

The EMBO journal ·Vol. 3 ·No. 11 ·1984-11-00 ·Pages 2445-52

Laski FA, Belagaje R, Hudziak RM, Capecchi MR, Norton GP, Palese P, RajBhandary UL, Sharp PA

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.

MeSH Terms
Amino Acid Sequence Animals Base Sequence Cell Line Chlorocebus aethiops Cloning, Molecular Codon DNA Restriction Enzymes Genes, Synthetic Kidney Mutation Nucleic Acid Conformation RNA, Transfer, Amino Acyl/genetics Simian virus 40/genetics Suppression, Genetic Transcription, Genetic Xenopus
Chemicals
Codon RNA, Transfer, Amino Acyl DNA Restriction Enzymes
Authors & Affiliations
8 authors, click to expand affiliations / ORCID
Laski F A
Belagaje R
Hudziak R M
Capecchi M R
Norton G P
Palese P
RajBhandary U L
Sharp P A
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Article Info
Journal
The EMBO journal
Abbr.
EMBO J
ISSN
0261-4189
Published
1984-11-00
Pages
2445-52
Language
English
Region
England
NLM ID
8208664
PMCID
PMC557710
Subset
IM
Grants
NIAID NIH HHS · AI-11823 · United States
NIGMS NIH HHS · GM17151 · United States
NIGMS NIH HHS · GM21168 · United States
External Links
PubMed source PMC Full Text View journal details
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