Functional assay of tRNA molecules transcribed from a purified gene

L. Corbo, G. Ciliberto, C. Traboni, R. Santamaria, F. Cimino, R. Cortese, F. Salvatore

Research output: Contribution to journalArticle

Abstract

Purified tRNA genes are expressed when microinjected into the nucleus of X.laevis oocytes. In this paper we describe a method to assay the capacity to be aminoacylated of the tRNA transcribed in the frog oocytes. The method exploits the radiochemical purity of the transcript and relies on the binding of aminoacyl-tRNA but not of uncharged tRNA to purified elongation factor EF-TU. We also present some preliminary results on several single point mutants of tRNAPro from Caenorhabditis elegans. We show that nucleotide 73 of tRNAPro can be substituted by any other nucleotide without loss of acceptor activity. A doublemutant, causing transition from G45G46 to A4546 has lost acceptor activity. Also inactive is a mutant carrying an insertion of a single base in the anticodon loop.

Original languageEnglish
Pages (from-to)7363-7372
Number of pages10
JournalNucleic Acids Research
Volume10
Issue number22
DOIs
Publication statusPublished - Nov 25 1982

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ASJC Scopus subject areas

  • Statistics, Probability and Uncertainty
  • Applied Mathematics
  • Health, Toxicology and Mutagenesis
  • Toxicology
  • Genetics(clinical)
  • Genetics

Cite this

Corbo, L., Ciliberto, G., Traboni, C., Santamaria, R., Cimino, F., Cortese, R., & Salvatore, F. (1982). Functional assay of tRNA molecules transcribed from a purified gene. Nucleic Acids Research, 10(22), 7363-7372. https://doi.org/10.1093/nar/10.22.7363