The first evidence for a plant tRNA(Lys)(CUU) gene is reported. This gene is found closely linked 400 bp upstream, and on the complementary strand, of a ZmFer2 ferritin gene in the maize nuclear genome. Southern blot analysis indicates that this tRNA(Lys) is a member of a multigene family. This gene does not contain any intron, and exhibits classical intragenic regulatory elements found in eukaryotic tRNA genes (A and B boxes). Moreover, 5' and 3'-flanking sequences display typical features found in nuclear encoded tRNAs. The deduced mature tRNA sequence is almost identical to the sequence of a cytoplasmic tRNA(Lys)(CUU) from wheat germ. The maize tRNA(Lys) gene is expressed in vivo in maize and in transgenic tobacco, as shown by RT-PCR analysis.