Worm Breeder's Gazette 10(1): 17
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.
On the right side of the cluster on Chromosome V are several genes which, when mutated, cause defective cuticles. These genes include vab-8, rol-4, cuticles are comprised predominantly of collagens, I wanted to determine whether any of these genes do in fact encode a collagen. Using radioactivity-labeled pcol-2, I probed restriction digests of overlapping cosmids contained within the myo-3 contig which had been mapped to this region. (See maps below). Two of these cosmids, C12A12 and ZC348, cross-hybridized to pcol-2 under low stringency conditions. Interestingly, the HindIII fragment of ZC348 had the same molecular weight (3.5 Kbp) as did the HindIII fragment of pcol-1. As a further test for identity, I probed double digests of both ZC348 and pcol-1 (HindIII/BamHI and Hind III/HinFI) with labeled pcol-1 under high stringency conditions. Identical restriction fragments were seen for both clones. The localization of col-1 to this region is a tremendous aid to our study of cuticle-defective mutants: col-1 has been sequenced (Kramer et al., 1982, Cell 30, 599) and its pattern of developmental expression has been determined (Cox & Hirsh, 1985, Mol. Cell. Biol. 5, 363). We are now using the col-1 gene as a probe to look for RFLPs in mutant DNAs. To study the function of the col-1 gene product we plan to isolate specific antibodies which can be used for biochemical and morphological analyses of wild-type and mutant cuticles. [See figure 1]