GENERATION OF A 3-MB PAC CONTIG SPANNING THE MIYOSHI MYOPATHY LIMB-GIRDLE MUSCULAR-DYSTROPHY (MM/LGMD2B) LOCUS ON CHROMOSOME 2P13/

Miyoshi myopathy (MM) and limb-girdle muscular dystrophy subtype 2B (L GMD2B) map to the same region on chromosome 2p13. To facilitate the cl oning of the defective gene causing these two diseases, we used a comb ination of chromosome walking and expressed sequence tag (EST) screeni ng and identified 864 P1-derived artificial chromosomes (PACs) whose i nserts map to the MM/LGMD2B candidate region and surrounding areas. Am ong them, 139 are from a chromosome 2-specific PAC library and 725 are from a total genomic PAC library. A 3-Mb contig spanning the candidat e region for MM/LGMD2B was assembled. This contig contains 200 PACs, 1 0 known genetic markers, 5 new polymorphic markers, 57 sequence tagged sites (STSs) generated from PAC end fragments, and 4 random STSs. In addition, we mapped 24 ESTs to this contig and excluded 37 ESTs from t he contig, thus eliminating them as candidate MM/LGMD2B genes. The hig h-resolution, sequence-ready PAC contig for the MM/LGMD2B region provi des a backbone for the identification of the disease gene(s) and for c larification of the relationship between the two diseases. (C) 1998 Ac ademic Press.