A 6-MB YAC CONTIG IN XP22.1-P22.2 SPANNING THE DXS69E, XE59, GLRA2, PIGA, GRPR, CALB3, AND PHKA2 GENES

We report the generation of an similar to 6-Mb contig of 70 overlapping yeast artificial chromosomes (YAC) covering the interval between DXS16 and DXS1229 in Xp22.1-p22.2. Within this region lie the genes for calbindin (CALB3), gastrin-releasing peptide receptor (GRPR), phosphatidyl-inositol glycan-class A protein (PIGA), glycine receptor alpha-2 (GLRA2), phosphorylase kinase alpha (PHKA2), XE59 (a gene escaping X chromosome inactivation), and DXS69E (71-7A). YACs were isolated initially from four libraries either by hybridization or using sequence tagged sites (STSs) for DXS16, DXS9, GLRA2, DXS207, DXS43, DXS1416, DXS1317, DXS1195, and DXS418. Additional STSs were obtained from the end fragments of the original YACs studied, thus allowing us to cover the contig with a series of 73 STSs, approximately 1 per 100 kb. YAC contig construction allowed the following locus order to be established: Xpter-DXS16-DXS69E-DXS414-XE59-DXS9-(GLRA2, DXS987)-(PIGA, DXS207)-DXS1053-DXS197-(GRPR, DXS43)-CALB3-DXS1416-DXS1317-DXS1195-DXS418-DXS257-(PHKA2, DXS999)-DXS443-DXS1229-Xcen. Restriction mapping of the DXS16-DXS43 interval predicted the existence of several CpG islands, suggesting the presence of other genes in the region. This work provides a starting point for further mapping and positional cloning of several X-linked disease genes. (C) 1995 Academic Press, Inc.