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Genetics, Vol 123, 337-348, Copyright © 1989
INVESTIGATIONS |
Physical and Functional Definition of the Drosophila Notch Locus by P Element Transformation
RGP. Ramos, B. G. Grimwade, K. A. Wharton, T. N. Scottgale and S. Artavanis-Tsakonas
Department of Biology, Yale University, New Haven, Connecticut 06520
Notch is a developmentally regulated locus which controls the differentiation of various Drosophila tissues, among them the embryonic nervous system. Molecular analysis has suggested that Notch is defined by an approximately 40-kb transcription unit which is spliced into a 10.2-kb mRNA composed of nine exonic regions and coding for a 2703-amino acid long transmembrane protein that shows homology to the mammalian epidermal growth factor. Here, we define the 5' end of the transcription unit and determine the sequences deleted in a Notch mutation involving the 5' nontranscribed region. Using a Notch cosmid vector we demonstrate by P element-mediated transformation that all sequences necessary for Notch function are confined in an approximately 40-kb long genomic region. cDNA sequences are used to construct a 15-kb ``minigene'' which lacks most, but not all, introns and its functionality is also tested by P element transformation. We show that, unlike the cosmid vector which is capable of rescuing completely all Notch mutations, only certain phenotypes can be rescued by the ``minigene.'' The functional implications of our findings are discussed.
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