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Identification of genes required for Drosophila eye development using a phenotypic enhancer-trap

Author:
Pignoni, F., Hu, B., Zipursky, S.L.
Source:
Proceedings of the National Academy of Sciences of the United States of America 1997 v.94 no.17 pp. 9220-9225
ISSN:
0027-8424
Subject:
ommatidia, insertional mutagenesis, photoreceptors, neurons, phenotype, mutants, reporter genes, biological development, Drosophila melanogaster, transposons
Abstract:
A novel method of P-element mutagenesis is described for the isolation of mutants affecting the development of the Drosophila compound eye. It exploits the interaction between the Bride of Sevenless (Boss) ligand and the Sevenless (Sev) receptor tyrosine kinase that triggers the formation of the UV-sensitive photoreceptor neuron, R7. Transposition of a boss cDNA transgene, in an otherwise boss mutant background, was used as a "phenotypic trap" in live flies to identify enhancers expressed during a narrow time window in eye development. Using a rapid behavioral screen, more than 400,000 flies were tested for restoration or R7. Some 1,800 R7-containing flies were identified. Among these, 21 independent insertions with expression of the boss reporter gene in the R8 cell were identified by a external eye morphology and staining with an antibody against Boss. Among 900 lines with expression of the boss reporter gene in multiple cells assessed for homozygous mutant phenotypes, insertions in the marbles, glass, gap1, and fasciclin II genes were isolated. This phenotypic enhancer-trap facilitates (i) the isolation of enhancer-traps with a specific expression pattern, and (ii) the recovery of mutants disrupting development of specific tissues. Because the temporal and tissue specificity of the phenotypic trap is dependent on the choice of the marker used, this approach can be extended to other tissues and develop mental stages.
Agid:
2356648