GENOMIC ORGANIZATION OF THE NEUROFIBROMATOSIS-1 GENE (NF1)

Neurofibromatosis 1 maps to chromosome band 17q11.2, and the NF1 locus has been partially characterized. Even though the full-length NF1 cDN A has been sequenced, the complete genomic structure of the NF1 gene h as not been elucidated, The 5' end of NF1 is embedded in a CpG island containing a NotI restriction site, and the remainder of the gene lies in the adjacent 350-kb NotI fragment. In our efforts to develop a com prehensive screen for NF1 mutations, we have isolated genomic DNA clon es that together harbor the entire NF1 cDNA sequence. We have identifi ed all intron-exon boundaries of the coding region and established tha t it is composed of 59 exons. Furthermore, we have defined the 3'-untr anslated region (3'-UTR) of the NF1 gene; it spans approximately 3.5 k b of genomic DNA sequence and is continuous with the stop codon. Oligo nucleotide primer pairs synthesized from exon-flanking DNA sequences w ere used in the polymerase chain reaction with cloned, chromosome 17-s pecific genomic DNA as template to amplify NF1 exons 1 through 27b and the exon containing the 3'-UTR separately. This information should be useful for implementing a comprehensive NF1 mutation screen using gen omic DNA as template. (C) 1995 Academic Press, Inc.