Worm Breeder's Gazette 11(2): 58

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 unc-73

Rob Steven and Joe Culotti

Unc-73 affects the longitudinal growth of certain neurons and the 
excretory cell as well as certain longitudinal cell migrations on the 
nematode epidermis (Hedgecock et al., Development 100, 365-382, 1987). 
We previously isolated unc-73(ev454) on strain RW7097 and identified 
a Tc1 on a 5 kb HindIII which cosegregated with this mutation in 3 
factor crosses (CSH abstracts, 1989, p.  60).  We have now cloned this 
5 kb Tc1-containing HindIII fragment, excised the Tc1 with EcoRV, and 
used the flanking sequence to probe Southern blots of DNAs from unc-73(
ev454) and from two independently derived spontaneous revertants of 
unc-73(ev454).  As expected, this probe hybridized to a 5 kb HindIII 
fragment on the unc-73(ev454) lane and to a 3.4 kb HindIII fragment on 
the revertant lanes.  No polymorphisms were observed in DNA from two 
EMS-induced alleles of unc-73 (e936 and rh40).  We have obtained 
approximately 250 nucleotides of sequence from both ends of the 5 kb 
HindIII fragment.  One of these ends comprises a GC rich open reading 
frame which fails to show any significant homology to anything in the 
NBRF database.  We have also identified about a dozen phages from an 
EMBL-3 N2 genomic library (Nishiwaki and Miwa) which hybridize to our 
Tc1-flanking probe, but we have not yet characterized any of these.  
We hope that these will allow Coulson and company to identify the unc-
73-containing contig.