Investigators' Abstract) The goal of this project is the construction of high resolution genetic and physical maps of the murine X chromosome from clotting factor 9 to Duchenne muscular dystrophy. This region contains one of the sites of chromosomal rearrangement between the human and murine X chromosomes as well as loci corresponding to several human disease genes. The region covers 10-12 cM or approximately 12-15 million base pairs (Mb). Recombinant backcross progeny from an interspecific mating of Mus domesticus and M. spretus will be typed for 8 anchor loci spanning the murine X chromosome and form a genetic mapping panel. 270 backcross progeny have been produced, and a total of 400 animals will be generated. DNA probes spaced 0.3 to 0.5 cM apart spanning the region from Cf-9 to Dmd will be identified. Existing human and murine probes which will be mapped include (a) random murine X """"""""linking clones"""""""" that map proximal to Pgk-1 and (b) human YAC (yeast artificial chromosome) clones from Xq27-Xq28. An adaptation of an efficient method to generate probes from human YACs is proposed to enable cross-species hybridization and mapping. Additional random murine X probes will be generated from mouse X:human hybrids using polymerase chain reaction (PCR) amplification of DNA between mouse repetitive sequences, a technique called interspersed repetitive sequence (IRS) PCR. Preliminary mapping of PCR-generated probes from a hybrid containing a fragment of the mouse X demonstrates the feasibility of the method. Additional deleted-X hybrids will be generated by X-irradiation of the mouse X:human hybrids. A physical map of the region will be compiled using the deleted X hybrids, pulsed field gel electrophoresis (PFGE), and alignment of overlapping YAC clones. Genetic and physical maps will be compared. Maps of the region will be compared with the corresponding region of the human X (Xq27-Xq28).

Agency
National Institute of Health (NIH)
Institute
National Human Genome Research Institute (NHGRI)
Type
Research Project (R01)
Project #
5R01HG000131-03
Application #
2208541
Study Section
Genome Study Section (GNM)
Project Start
1991-09-30
Project End
1995-08-31
Budget Start
1993-09-01
Budget End
1995-08-31
Support Year
3
Fiscal Year
1993
Total Cost
Indirect Cost
Name
Baylor College of Medicine
Department
Genetics
Type
Schools of Medicine
DUNS #
074615394
City
Houston
State
TX
Country
United States
Zip Code
77030
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Pasteris, N G; de Gouyon, B; Cadle, A B et al. (1995) Cloning and regional localization of the mouse faciogenital dysplasia (Fgd1) gene. Mamm Genome 6:658-61
Rogner, U C; Kioschis, P; Wilke, K et al. (1994) A YAC clone map spanning 7.5 megabases of human chromosome band Xq28. Hum Mol Genet 3:2137-46
Chatterjee, A; Faust, C J; Herman, G E (1993) Genetic and physical mapping of the biglycan gene on the mouse X chromosome. Mamm Genome 4:33-6