Previous experiments have demonstrated that the information suppressors
sup-5 and
sup-7 are tRNA(trp) amber suppressors (Cell 33, 575-583; P.N.A.S. 81, 6784-6788 our unpublished results). By Southern analysis using a tRNA(Trp) gene as a probe there appears to be twelve members in the tRNA(Trp) family (P.N.A.S. 81, 6784-6788). We have now cloned all twelve members of this tRNA family and are presently sequencing each member. Part of our studies of this gene family are aimed at trying to isolate suppressor alleles of each member. From reversion studies of
unc-13(
e1091); 64) using nitroso-guanidine as a mutagen we have isolated a new amber suppressor mutation,
st354. This new mutation is a weaker suppressor than either
sup-5 or
sup-7. Altering a tRNA(Trp) to an amber suppressor creates an XbaI site in the anticodon loop. By Southern analysis we have demonstrated that
st354 has a new XbaI site in a 10kb XbaI fragment which contains a tRNA(Trp) family member. Genetic mapping studies reveal that
st354 is loosely linked to dpy- 13 on chromosome IV. By 3-factor mapping we have placed it to the right of
dpy-4. This data is shown below: [See Figure 1] From a 2-factor cross we have determined that
st354 is about 0.6 map units right of
ipy-4. This tRNA(Trp) gene is at present the most distal marker on the right half of chromosome IV.