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Nucleic Acids Research, 1984, Vol. 12, No. 14 5737-5755
© 1984

Articles

Accurate transcription of homologous 5S rRNA and tRNA genes and spllcing of tRNA in vitro by soluble extracts of Neurospora

Brett M. Tyler and Norman H. Giles

Department of Molecular and Population Genetics, University of Georgia Athens, GA 30602, USA

Received February 21, 1984. Revised June 18, 1984. Accepted June 18, 1984.

We have developed soluble extracts from Neurospora crassa capable of accurately and efficiently transcribing homologous 5S rRNA and tRNA genes. The extracts also appear to quantitatively end-process and splice the primary tRNA transcripts. Although the extracts could not transcribe a heterologous (yeast) 5S rRNA gene, they did transcribe a yeast tRNALeu gene and slowly process the transcripts. In addition, we have developed a novel strategy for rapidly sequencing uniformly labelled RNAs using base-specific ribonucleases. We have used this procedure to verify the identity of the in vitro tran scripts and processing products.


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