THE RNA SUBUNIT OF RIBONUCLEASE-P FROM THE ZEBRAFISH, DANIO-RERIO

A simple strategy has been devised to identify the gene encoding the R NA subunit of RNase P from the zebrafish, Danio rerio. The sequence ob tained by amplification of genomic DNA with primers based on sequences common to two other vertebrates was confirmed by reverse transcriptio n and amplification of RNA from a partially purified preparation of th e holoenzyme. The 5' and 3' ends were determined by cyclizing the RNA, followed by reverse transcription and sequencing across the ligated R NA junction, The zebrafish sequence is 63% identical to that of Xenopu s laevis nuclear RNase P RNA and 69% identical to the human RNase P RN A, A consensus secondary structure was constructed based on these nucl eotide identities and on the many compensatory base changes in several regions among these three RNAs. The strategy used to obtain the zebra fish sequence should be useful in deriving analogous gene sequences fr om diverse classes of eukaryotes.