Bacteriophage T5 transfer RNA. Isolation and characterization of tRNA species and refinement of the tRNA gene map.

Previous studies from this laboratory have provided a high resolution map for 16 tRNA genes located on the continuous heavy DNA strand of bacteriophage T5 DNA (Chen, M.-J., Locker, J., and Weiss, S.B. (1976) J. Biol. Chem. 251, 536-547). All of the T5 tRNA genes were located in three clusters within the DNA C segment, except for tRNAArg, which mapped on the left end of the DNA D segment (Desai, S.M., Hunt, C., Locker, J., and Weiss, S.B. (1978) J. Biol. Chem. 253, 6544-6550). In this report, we present evidence for the presence of eight additional T5 tRNA species, five of which are located in two new loci within the DNA C segment. We also describe a two-dimensional gel electrophoresis system for the separation and isolation of T5 tRNA species from crude infected RNA preparations. The gel electrophoresis system separates tRNA isoacceptors specific for different amino acids; evidence is presented that the isoacceptors for isoleucine, histidine, and serine are coded by different T5 genes.