Worm Breeder's Gazette 11(5): 19
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.
We are interested in studying the molecular genetics of sensory neuron development. Two genes previously implicated in sensilla development, particularly in the development of sensory cilia, are daf- 10(IV) and osm-1(X) (Perkins et. al. 1986 Dev. Biol. 117, 456487). Mutant animals have a variety of ciliary defects. However, the major defect appears to be abnormally short axonemal projections and (at least for osm-1) an assembly of ectopic doublet microtubules which constitute cilia-like posterior projections. These animals have various behavioral abnormalities which presumably result from a failure of the sensory cilia to project into the environment (i.e. daf, osm, che etc.). To understand how these genes function, we have begun a molecular characterization of the two loci. We obtained from Don Riddle's lab alleles osm-1(m530), 8), and daf-10(m534), which were generated in RW7097. Spontaneous revertants were identified and, they and the mutants from which they were derived were outcrossed. DNA samples from outcrossed mutants, revertants, and recombinants were isolated for Southern analysis. For each gene, a single Tc1- containing EcoRI fragment which co-segregated with the mutant phenotype was identified. These EcoRI fragments were cloned and their identities verified. DNA fragments flanking the Tc1 elements were used to screen a genomic library provided by Chris Link. From these experiments we identified three overlapping lambda clones for osm-1 and a single lambda clone which hybridized to the daf-10 probe. The osm-1 and daf-10 lambda clones were sent to John Sulston and Alan Coulson at the MRC for assignment on the physical map. Both genes landed in large contigs. The daf-10 clone was placed within approximately 20kb and to the right of fem-3, and the osm-1 clone was placed within about 100kb and to the right of mtl-1. In addition fragments of the genomic clones were used to identify hybridizing cDNA clones from a library provided by Stuart Kim. The osm-1 and daf-10 probes each identified cDNAs. We have begun sequencing these clones and using them as probes for northern analyses.