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Nucleic Acids Research, 1980, Vol. 8, No. 22 5169-5178
© 1980

MOLECULAR BIOLOGY

Nonsense suppression in eukaryotes: the use of the Xenopus oocyte as an in vivo assay system

Mariann Bienz*, Eric Kubli*, Kohli Jürg+, Suzanne de Henau{dagger} and Henri Grosjean{dagger}

*Zoological Institute, University of Zürich Winterthurerstrasse 190, CH-8057 Zürich +Institute of Microbiology, University of Berne Altenbergrain 21, CH-3013, Switzerland {dagger}Laboratoire de Chimie Biologique, Université Bruxelles 67 Rue des Chevaux, B-1640 Bruxelles, Belgium

Received October 7, 1980. Amber, ochre, and opal nonsense suppressor tRNAs isolated from yeast were injected into Xenopus laevis oocytes together with purified mRNAs (globin mRNA from rabbit, tobacco mosaic virus-RNA). Yeast opal suppressor tRNA is able to read the UGA stop codon of the rabbit ß-globin mRNA, thus producing a readthrough protein. A large readthrough product is also obtained upon coinjection of yeast amber or ochre suppressor tRNA with TMV-RNA. The amount of readthrough product is dependent on the amount of injected suppressor tRNA. The suppression of the terminator codon of TMV-RNA is not susceptible to Mg++ concentration or polyamine addition. Therefore, the Xenopus laevis oocyte provides a simple, sensitive, and well buffered in vivo screening system for all three types of eukaryotic nonsense suppressor tRNAs.


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