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Drosophila NF1 clones were isolated by screening a Canton-S λFixII genomic phage library (Stratagene) in 25% formamide at 37°C with a probe representing the COOH-terminal 1598 amino acids of human NF1. All clones identified represented the same locus. A 13,295-bp segment representing the entire gene was sequenced. A set of 32 overlapping cDNAs was isolated from eye disc and mixed stage embryo libraries and used to determine 9750 bp of overlapping cDNA sequence. The genomic and cDNA sequences differ in multiple locations, but none of these polymorphisms affects the sequence of the encoded protein. The Gen-Bank accession numbers for the genomic and alternatively spliced cDNA sequences are L26500, L26501, and L26502.
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2 males of genotype w; P[lacZw]/TM3 or w; P[lacZw]/TM6B were crossed to w; TM3/TM6B virgin females to establish lines with stable P-element integrations. The red-eyed progeny of this cross were analyzed in pools by inverse polymerase chain reaction [B. Dalby, A. J. Pereira, L. S. B. Goldstein, Genetics 139, 757 (1995)].
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P1 or f ; bld[f+]/+ were analyzed.
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We thank T. Orr-Weaver and F. Shamanski for help with cytogenetic mapping; C. Ngwu for assistance with monoclonal antibody production; G. Bollag for GAP and human NF1 fusion proteins; S. Artavanis-Tsakonas, J. de Celis, S. Harrison, D. Kalderon, T. Laverty, G. Rubin, and G. Struhl for fly stocks; L. Zipursky for stocks and 96F DNA clones; and S. Brill, J. Settleman, and S. van den Heuvel for critical reading of the manuscript. Supported by a grant from the U.S. Army Medical Research and Materiel Command to I.K.H. and A.B., a grant from NF Inc. (Massachusetts Bay Chapter) to J.F.G., NIH grants to J.F.G. (NS22229), I.K.H. (NS36084), Y.Z. (NS34779), and A.B. (NS31747), and a postdoctoral fellowship from the Medical Research Council of Canada to G.E.H.
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