International Mammalian Genome Society

The 14th International Mouse Genome Conference (2000)


A24. Comparative Sequencing of the Mouse X Chromosome: F8 to Dmd

Bate R1, Straw R2, Mallon A-M1, Botcherby MRM2, Strivens MA1, Weston P2, Hunter C2, Gilbert M2, Goodall K2, Kerry G2, Greystrong J.S.2, Clarke D2, Rhodes M2, Campbell R2, Brown SDM1, Denny P1
1 MRC UK Mouse Genome Centre and Mammalian Genetics Unit, Harwell, Oxon, UK
2 MRC Human Genome Mapping Project Resource Centre, Hinxton, Cambridge, CB10 1SB, UK

The 2-3 Mb region that spans from F8 (Factor VIII) to Dmd (Duchenne muscular dystrophy) on the mouse X chromosome crosses two evolutionary breakpoints with F8 and Dmd residing on human Xq28 and Xp21 respectively. Recent work (1) has also placed Tbl1 (Transducin beta-like 1) between F8 and Dmd, which maps to human Xp22.3. Work is currently focused on generating a sequence-ready map between F8 and Dmd, with particular interest in identifying and ultimately sequencing across the evolutionary breakpoints.

A genetic map has been created across the complete region using the EUCIB resource. Due to the paucity of available genes within the F8 to Dmd region, microsatellites and STSs are being used to anchor clones to genetic, radiation hybrid and physical maps. A PCR based approach has been employed to screen pools generated from the RPCI21 and RPCI23 PAC & BAC libraries, firstly with MIT microsatellite markers, then with primers designed from clone end-sequence data. To complement this approach, high-density filters from the same libraries have been screened by hybridization using labelled probes generated from short over-lapping oligos (overgos). A physical contig has been generated around Tbl1 and further contigs around proximal and distal MIT microsatellite markers. Significant portions of this region are being sequenced to target the location of the breakpoints.

We will present progress in map construction, definition of the evolutionary breakpoints and sequence analysis.

1. C. M. Disteche, M. B. Dinulos, M. T. Bassi, R. W. Elliott and E. I. Rugarli. Mamm. Genome 9, 1062-1064 (1998)


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