The nucleotide sequence of a cloned Drosophila arginine tRNA gene and its in vitro transcription in Xenopus germinal vesicle extracts

Abstract

The DNA sequence of a cloned gene specifying Drosophila tRNA2Arg has been determined. Transcription of this gene in extracts from Xenopus germinal vesicles yielded a precursor RNA containing a seven-nucleotide leader sequence at the 5' side of the mature tRNA with pppGp as the 5'-terminal nucleotide. Transcription continues for at least eight nucleotides past the 3' terminus of the coding region of the gene and terminates over a stretch of seven T residues. The isolation of the primary transcript was facilitated by incorporation of 5'-(gamma-S)GTP into the 5' terminus of the precursor RNA and by its retention on mercury-agarose. The Drosophila tRNAArg gene does not contain an intervening sequence nor the C-C-A sequence corresponding to the 3' terminus of the mature tRNA. The nucleotide sequence of Drosophila tRNAArg formed in the in vitro system differs only in four positions from that of mouse tRNAArg.