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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.
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