Transcription of eukaryotic genes with impaired internal promoters: the use of a yeast tRNA gene as promoter

• Published in: Journal of biotechnology Vol. 21; no. 3; pp. 289 - 293
• Main Authors: Otter, Charlotta A., Straby, Kerstin B.
• Format: Journal Article
• Language: English
• Published: Lausanne Elsevier B.V 1991; Amsterdam Elsevier; New York, NY
• Subjects: Artificial promoter; artificial promoters; Biological and medical sciences; Biotechnology; Dimeric transcripts; Fundamental and applied biological sciences. Psychology; gene expression; genes; Genes, Fungal; Maturation; messenger RNA; Molecular and cellular biology; Molecular genetics; Plasmid constructions; Plasmids; Processing; promoter regions; Promoter Regions, Genetic; RNA editing; RNA, Transfer - genetics; Saccharomyces cerevisiae; Saccharomyces cerevisiae - genetics; transcription (genetics); Transcription, Genetic; Transcription. Transcription factor. Splicing. Rna processing; transfer RNA; tRNA gene pairs; Dimeric transcripts; Artificial promoter; Maturation; Plasmid constructions; tRNA gene pairs; Processing; Yeast; Molecular processing; Transcription; Transcription promoter; Fungi; Plasmid; Messenger RNA; Arginine; Gene; t-RNA; Ascomycetes; Aspartic acid; Saccharomyces cerevisiae; Thallophyta
• ISSN: 0168-1656; 1873-4863
• DOI: 10.1016/0168-1656(91)90049-2

Plasmid construction carrying promoters that allow eukaryotic genes with nonfunctional inherent promoters to be transcribed both in vitro and in vivo in eukaryotic systems is a useful and in certain cases necessary tool in biotechnology. Promoter and UAS sequences of Pol I and II, transcribing rRNA and structural genes respectively, are not always easily identifiable and even if so, not always possible to use in practice. This report summarizes results of using a tRNA gene with internal promoters as an "artificial" promoter for any DNA linked behind the tRNA gene via a short spacer. In vitro the reaction is catalyzed by a yeast pol III transcription and processing system. Thus both primary and processed RNA can be isolated for different purposes. In vivo the genes can be expressed in yeast cells, allowing the observation of biological effects of mutations, and in oocytes.