Worm Breeder's Gazette 9(3): 65
These abstracts should not be cited in bibliographies. Material contained herein should be treated as personal communication and should be cited as such only with the consent of the author.
Using a Xenopus laevis initiator methionine tRNA (tRNA meti) gene as a probe, we have screened a C. library ( constructed by T. Snutch) to isolate the nematode tRNA meti genes. Eighteen positives were isolated and plaque purified. Analysis of these clones by Southern transfer and restriction digestion shows that the eighteen positives fall into four different groups of phages. Southern blots of EcoR1 restricted C. eight bands hybridizing to the Xenopus probe. These bands are single copy and are dispersed throughout the genome. Detailed restriction maps of the four phages isolated confirm that they come from different genomic locations. A small piece of DNA containing the putative gene from each phage was subcloned into pUC19 to allow further characterization. We have sequenced two of the tRNA meti genes and both have identical coding sequences. We are now in the process of sequencing the other two genes. These subclones were also used to program our in vitro transcription system. All four clones produced both a precursor and a processed, mature tRNA product. We also plan to use the flanking phage fragments as templates in the nematode transcription system to look for the presence of any other pol 111 genes lying in close proximity to the cloned tRNA meti genes.