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.

Molecular Cloning of dpy-5

Joe Babity and Ann Rose

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.