Worm Breeder's Gazette 10(3): 53
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.
The molecular cloning of dpy-5(I) was initiated by genomic blot analysis of dpy-5(s1300), a spontaneous mutation isolated in an N2/BO hybrid strain by R. Rosenbluth. A novel 2.7 kb Tc1-hybridizing band was present in s1300 and cosegregated with the Dpy-5 phenotype. This Tc1 was shown to map between unc-38 and unc-87 through the construction of an unc-38 00) utant. Two additional dpy-5 strains and two non-Dpy revertants were analyzed: dpy-5(mn303) and a revertant kindly provided by Bob Herman and dpy-5(h14) and a revertant from our lab. DNA from both the Tc1-induced dpy-5(mn303) and the BO dpy-5(h14) contain the novel 2.7 kb Tc1-hybridizing band. Revertants of both mutations do not have this Tc1. Using unc-38 00) onstructed a lambda gt10 library of EcoRI fragments ranging in size from approximately 2500 bp to 2800 bp. This library contained three Tc1's, one of which is the novel 2.7 kb Tc1. DNA flanking the Tc1 was used as a probe in genomic blots of different Dpy-5 strains. In DNA from dpy-5(h14) and dpy-5(mn303), a 2.7 kb band is present, whereas revertants of these alleles contain 1.1 kb hybridizing bands. An additional Tc1-induced dpy-5, 76), kindly provided by Don Riddle), has the 1.1 kb hybridizing band. This may mean that the dpy-5 gene extends into an adjacent EcoRI fragment. Approximately 600 bp have been sequenced from DNA flanking the novel Tc1. As yet no collagen-like homology has been detected as was reported for the dpy-13 sequence (N. von Mende, D. Mck. Bird, P. S. Albert and D. L. Riddle, The Worm Breeder's Gazette, Vol. 10, No. 1). A genomic phage has been isolated by homology to putative dpy-5 sequences. This phage has been sent to John Sulston and Alan Coulson in Cambridge, England.