Studies on the amount and location of the tRNA and 5-S rRNA genes in Tetrahymena pyriformis GL.

The amount and location of tRNA and 5-S rRNA genes in the macronucleus of Tetrahymena pyriformis GL was investigated by DNA-RNA hybridization. Hybridization of 32P-labelled tRNA in excess of unlabelled rRNA (25-S + 17-S + 5-S) showed that at saturation 0.021% of the macronuclear DNA was complementary to tRNA. Hybridization of 32P-labelled 5-S rRNA in excess of unlabelled 25-S + 17-S rRNA and tRNA showed a saturation value of 0.017%. In contrast to the 25-S + 17-S rRNA genes, which are found in DNA of high bouyant density, tRNA and 5-S rRNA genes were distributed evenly throughout the main peak observed when bulk macronuclear DNA was fractionated by density centrifugation in CsCl gradients. Sucrose gradient analyses of total macronuclear DNA showed that tRNA and 5-S rRNA genes were found in DNA of all size classes but a significant enrichment in the slowly sedimenting DNA fraction was observed. Saturation hybridization of 5-S rRNA to purified rDNA showed that rDNA did not contain any 5-S rRNA genes.