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3’ Terminal Processing of Precursor tRNA Transcribed From a Drosophila Melanogaster Histidine Gene in a Cell-Free System
Fulginiti, James P.
Fulginiti, James P.
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1987-01-01
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Abstract
Transfer RNA biosynthesis is a complex process which includes trimmings at the 5' and 3' termini and nucleotide modification of the initial tRNA precursor. This research involves the detention and isolation of a 3' endonucleolytic activity from Schizosaccharomyces pombe.
tRNA precursors are obtained from a cell-free transcription system using (i) a Drosophila tRNA-histidine gene which contains a 35 base pair trailer sequence at its 3' terminus and (ii) a crude yeast enzyme extract which can faithfully transcribe the gene and process the precursor transcripts. Transcription products are separated by means of polyacryfamide gel electrophoresis visualized by autoradiography, and eluted from the gel. The tRNA precursors are then incubated with a Sc. pombe extract, electrophoresed and autoradiographed. The intact 35 base pair trailer sequence will serve as an indicator of the presence of the 3' endonuclease.