Worm Breeder's Gazette 7(2): 16
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.
In the last newsletter, we described a selection of lethal amber mutations linked to dpy-10, using sup-7. We have revised this procedure slightly to take advantage of the balancer chromosome C1. Our current procedure is to mutagenize unc-4(e120); ) hermaphrodites and mate them with sup-7 males. Neither marker is suppressed by sup-7. From the F1 progeny, we pick wild-type animals and score their F2 progeny. Broods that have fertile Uncs and Dpys, but no fertile Unc Dpys are candidates for an amber lethal linked to unc-4. We have been outcrossing such candidates with C1/unc-4 males to establish stocks of genotype C/unc-4 am, which segregate am am mutant homozygotes. We also have been establishing stocks of genotype C1/unc-4 am; sup-7/sup-7. This is readily distinguished from the former stock not only by the suppressible lethal, but also by the suppression of unc-52(e669), one of the markers on the C1 balancer. Thus far, we have isolated more than thirty candidates, and have outcrossed about half. From these, two separate amber mutants have been maintained in both suppressed and nonsuppressed stocks. The balancer makes stock maintenance and mutant complementation much easier. We hope to identify amber mutations of genes known only by temperature-sensitive alleles, as well as new essential genes.