Worm Breeder's Gazette 11(2): 60

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The Cloning of lin-40

R.C. Johnsen and D.L. Baillie

Figure 1

From a Tc1 mutagenesis screen in the nT1(IV,V) balanced region using 
mut-4 RW7037, six lethal mutations that fail to complement lin-40 were 
recovered (Clark et al.  In Press Genome).  We have extracted DNA from 
the six strains containing lin-40 Tc1 mutations.  We digested the DNA 
with EcoRI, ran it on a gel, transfered the DNA to a filter and 
hybridized nick translated Tc1 DNA to it.  Three strains showed a 
common new Tc1 band.  For one allele (s1351) we recovered the new band,
isolated DNA flanking the Tc1 sequence and used the flanking sequence 
to screen T.  Snutch's Charon IV library.  From the library we 
isolated a phage (BC#S1001).  DNA from BC#S1001 was mapped to LGV(left)
on the cosmid map by J.  Sulston and A.  Coulson (see figure).  The 
location is surprising in that BC#S1001 maps very close to unc-60 
molecularly (unc-60 is in F53E2 pers.  comm.  M.  Wakarchuk) but 
genetically lin-40 is fairly far away (roughly 9 m.u.) to the right of 
unc-60.  We are attempting to rescue lin-40 by microinjection using 
appropriate cosmids.  We are also screening for revertants of some of 
the Tc1 alleles of lin-40.  This research has been supported by an MRC 
studentship to RCJ and NSERC and MDA (Canada) grants to DLB.
[See Figure 1]

Figure 1