Characterization of transcription and processing from plasmids that use polIII and a yeast tRNA gene as promoter to transcribe promoterdeficient downstream DNA

• Published in: Biochimica et biophysica acta. Gene structure and expression Vol. 1131; no. 1; pp. 62 - 68
• Main Authors: Otter, Charlotta A., Edqvist, Johan, Stråby, Kerstin B.
• Format: Journal Article
• Language: English
• Published: Amsterdam Elsevier B.V 07.05.1992; Elsevier
• Subjects: Biological and medical sciences; Biotechnology; Fundamental and applied biological sciences. Psychology; Genetic engineering; Genetic technics; Methods. Procedures. Technologies; polIII transcription; RNA processing; tRNA gene pair; Vectors (cloning, transfer, expression). Insertion sequences and transposons; Yeast; tRNA gene pair; Yeast; RNA processing; polIII transcription; Enzyme; Transcription promoter; Functional deficit; Recombinant DNA; Fungi; Gene; t-RNA; RNA polymerase 3; Helmintha; Ascomycetes; Nemathelminthia; Genetic engineering; Invertebrata; Nematoda; Saccharomyces cerevisiae; Thallophyta
• ISSN: 0167-4781; 1879-2634
• DOI: 10.1016/0167-4781(92)90099-L

Transfer RNA (tRNA) with mutations affecting the internal promoters and thereby causing them to be nontranscribable by polIII (exemplified, e.g., by nematode tDNA Pro with large insertions between the two internal promoters) could be transcribed by polIII both in vitro (yeast) and in vivo (oocytes) when cloned behind a yeast tRNA Arg gene. PolIII initiated RNA synthesis could also procede into a downstream structural gene normally read by polII. The resultant yeast-nematode dimeric tRNA linked in front of mRNA sequences was recognized by processing enzymes to give mature tRNA. Thus, a yeast tRNA gene preceded by its 5′ flank can function as a promoter for polIII transcription of any DNA, also of DNA coding for genes that otherwise could not have been expressed either in vitro or in vivo.