The nucleotide sequences of tobacco chloroplast genes for tRNAGly (UCC) and tRNAArg (UCU) have been determined. The tRNAGly gene has a 691-base-pair intron located in the D stem while the tRNAArg gene does not have any intron. The tRNAGly and tRNAArg genes are encoded on the same strand and separated by a 169-base-pair spacer. The tRNAGly gene is transcribed as a 900-base precursor RNA molecule. The tRNAGly and tRNAArg deduced from the DNA sequences show 84% and 55% sequence homologies with Escherichia coli tRNAGly (UCC) and phage T4 tRNAArg (UCU), respectively.