Isolation of DNA markers from a region between incontinentia pigmenti 1 (IP1) X-chromosomal translocation breakpoints by a comparative PCR analysis of a radiation hybrid subclone mapping panel.
A strategy based on the use of human-specific interspersed repetitive sequence (IRS)-PCR amplification was used to isolate regional DNA markers in the vicinity of the incontinentia pigmenti 1 (IP1) locus. A radiation hybrid (RH) resulting from a fusion of an irradiated X-only somatic cell hybrid (C12D) and a thymidine kinase deficient (TK-) hamster cell line (a23) was identified as containing multiple X chromosome fragments, including DNA markers spanning IP1 X-chromosomal translocation breakpoints within region Xp11.21. From this RH, a panel of subclones was constructed and analyzed by IRS-PCR amplification to (a) identify subclones containing a reduced number of X chromosome fragments spanning the IP1 breakpoints and (b) construct a mapping panel to assist in identifying regional DNA markers in the vicinity of the IP1 locus. By using this strategy, we have isolated three different IRS-PCR amplification products that map to a region between IP1 X chromosome translocation breakpoints. A total of nine DNA sequences have now been mapped to this region; using these DNA markers for PFGE analyses, we obtained a probe order DXS14-DXS422-MTHFDL1-DXS705. These DNA markers provide a starting point for identifying overlapping genomic sequences spanning the IP1 translocation breakpoints; the availability of IP1 translocation breakpoints should assist the molecular analysis of this locus.
Gorski
J L
JL
Department of Pediatrics, University of Michigan, Ann Arbor 48109.
Burright
E N
EN
Reyner
E L
EL
Goodfellow
P N
PN
Burgess
D L
DL
D000818:Animals; D015139:Blotting, Southern; D002874:Chromosome Mapping; D003001:Cloning, Molecular; D006224:Cricetinae; D004247:DNA; D016521:Electrophoresis, Gel, Pulsed-Field; D005819:Genetic Markers; D006801:Humans; D006822:Hybrid Cells; D007184:Incontinentia Pigmenti / Q000235:genetics*; D051379:Mice; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D011831:Radiation Genetics; D012091:Repetitive Sequences, Nucleic Acid; D014178:Translocation, Genetic; D014960:X Chromosome
D005819:Genetic Markers; D004247:DNA