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Studies on Arginyl Transfer Ribonucleic Acid in Herpes Virus Infected Baby Hamster Kidney Cells

Institute of Virology, University of Glasgow, Glasgow, W. 2, Scotland

Previous work, (Subak-Sharpe & Hay, 1965, Subak-Sharpe, Shepherd & Hay, 1966), depending on the hybridization of [³²P]4s RNA from herpes virus infected cells to herpes DNA and analysis of the aminoacyloligonucleotide fragments produced after RNase T 1 digestion of [¹⁴C]- and [³H]-arginyl tRNA, suggested the presence, in infected cells of a herpes-specified arginyl tRNA. The present study indicates that these results were due to the presence of (a) radioactive RNA other than tRNA in the [³²P] 4sRNA, and (b) radioactive contaminants in the [³H]-arginine used to aminoacylate the tRNA. Further purification of the aminoacyl tRNA removes these [³H]-arginine contaminants. Protection of the aminoacyl tRNA ester bond by N-acetylation improved the sensitivity of the aminoacyloligonucleotide analyses and permitted tRNA-specific molecular hybridization. Using these techniques, no herpes-specified arginyl tRNA was detected in cells 7 to 9 hr post infection.

^* Members of the Medical Research Council Virology Unit.

Received 7 May 1971; accepted 11 August 1971.