A proposal is made to analyze, by genetic and molecular techniques, a human gene whose expression is essential for normal neuroectodermal development. The research undertaken in this proposal will be directed towards the analysis of a human developmental gene, incontinentia pigmenti (IP). IP is an X-linked dominant, hemizygote lethal disorder. IP carrier females are identifiable and present with characteristic neuroectodermal developmental abnormalities. The IP locus has been tentatively localized to Xp11.21. This proposal provides for analysis of the IP locus by linkage analysis using collected genomic DNA samples from multigenerational IP families and characterized human X chromosomal probes capable of detecting restriction fragment length polymorphisms (RFLP's). An assemble collection of somatic cell hybrids containing rearranged X chromosomes will be used to isolated and map cloned X chromosomal fragments capable of detecting RFLP's, to the IP region. We propose to molecularly delineate the IP locus by identifying breakpoints of rearranged X chromosomes associated with the IP phenotype. Constructed somatic cell hybrids containing IP X/autosomal translocations will be used to identify X chromosomal overlaps, deletions, or breakpoint heterogeneity among the IP translocations by Southern blot analysis and pulse field gradient electrophoretic restriction mapping techniques. X chromosomal DNA fragments from the delineated Ip locus will be cloned. Unique, highly conserved regions from the IP locus will provide DNA fragment candidates for the isolation of the IP gene. These analyses will provide information to further delineate the genetic and chromosomal localization of a human developmental gene and provide the necessary starting point for cloning the IP gene. A more thorough understanding of neuroectodermal development, human developmental processes, genes and gen products necessary for normal completion of developmental process, and mutations that disrupt these processes should result from these studies.

Agency
National Institute of Health (NIH)
Institute
Eunice Kennedy Shriver National Institute of Child Health & Human Development (NICHD)
Type
Research Project (R01)
Project #
5R01HD023768-02
Application #
3324014
Study Section
Mammalian Genetics Study Section (MGN)
Project Start
1988-02-01
Project End
1991-01-31
Budget Start
1989-02-01
Budget End
1990-01-31
Support Year
2
Fiscal Year
1989
Total Cost
Indirect Cost
Name
University of Michigan Ann Arbor
Department
Type
Schools of Medicine
DUNS #
791277940
City
Ann Arbor
State
MI
Country
United States
Zip Code
48109
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Burright, E N; Gorski, J L (1993) Interspersed repetitive sequence (IRS)-PCR amplification of pulsed-field gel fractionated DNA to derive markers from the incontinentia pigmenti 1 (IP1) locus. Nucleic Acids Res 21:175-6
Gorski, J L; Burright, E N (1993) The molecular genetics of incontinentia pigmenti. Semin Dermatol 12:255-65
Pasteris, N G; Bialecki, M D; Gorski, J L (1993) YAC subclone contig assembly by serial interspersed repetitive sequence (IRS)-PCR product hybridizations. Nucleic Acids Res 21:5275-6
Gorski, J L; Boehnke, M; Reyner, E L et al. (1992) A radiation hybrid map of the proximal short arm of the human X chromosome spanning incontinentia pigmenti 1 (IP1) translocation breakpoints. Genomics 14:657-65
Gorski, J L; Burright, E N; Reyner, E L et al. (1992) 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. Genomics 14:649-56
Cotter, P D; Willard, H F; Gorski, J L et al. (1992) Assignment of human erythroid delta-aminolevulinate synthase (ALAS2) to a distal subregion of band Xp11.21 by PCR analysis of somatic cell hybrids containing X;autosome translocations. Genomics 13:211-2
Burright, E N; Reyner, E L; Gorski, J L (1991) Human-specific amplification of radiation hybrid DNA fractionated by pulsed-field gel electrophoresis. Nucleic Acids Res 19:401-2
Gorski, J L; Burright, E N; Harnden, C E et al. (1991) Localization of DNA sequences to a region within Xp11.21 between incontinentia pigmenti (IP1) X-chromosomal translocation breakpoints. Am J Hum Genet 48:53-64
Gorski, J L (1991) Father-to-daughter transmission of focal dermal hypoplasia associated with nonrandom X-inactivation: support for X-linked inheritance and paternal X chromosome mosaicism. Am J Med Genet 40:332-7

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