Abstract
X-linked mutant alleles associated with prenatal male lethality are difficult to analyze because only heterozygous females are readily available for study. Genomic analysis of the mutant allele is facilitated by the construction of somatic cell hybrids because this enables the segregation of the X Chromosomes (Chrs) that carry the mutant and wild-type alleles. We describe here a method that ensures that the X Chr carrying the mutant allele is retained in somatic cell hybrids in an active selectable state. This is achieved by mating heterozygous females to males that carry a mutation at the hypoxanthine phosphoribosyl transferase (Hprt) locus. The resultant F, females are compound heterozygotes, and when cells from these females are fused to HPRT-Chinese hamster cells and subjected to selection in HAT medium, the only survivors are those hybrid cells that retain an active X Chr carrying the mutant allele together with the wild-type Hprt allele. We use hybrids constructed by this method to demonstrate that there are no gross deletions or genomic rearrangements present in three mottled alleles associated with prenatal male lethality.
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References
Blair, H.J., Boyd, Y. (1994). Positioning of 14 simple sequence repeat loci relative to markers on the gene-based map of the mouse X chromosome. Mouse Genome 92, 127–129.
Chelly, J., Turner, Z., Tonnesen, T., Petterson, A., Ishikawa-Brush, Y., Tommerup, N., Horn, N., Monaco, A.P. (1993). Isolation of a candidate gene for Menke’; disease that encodes a heavy metal binding protein. Nature Genet. 1, 132–136.
Danks, D.M. (1977). Copper transport and utilisation in Menke’; syndrome and in mottled mice. Inorg. Perspect. Biol. Med. 1, 73–100.
Das, S., Levinson, B., Vulpe, C, Whitney, S., Gitschier, J., Packman, S. (1995). Similar splicing mutations of the Menkes/mottled copper-transporting ATPase gene in occipital horn syndrome and the blotchy mouse. Am. J. Hum. Genet. 56, 570–576.
Dierick, H.A., Ambrosini, L., Spencer, J., Glover, T.W., Mercer, J.F. (1995). Molecular structure of the Menke’; disease gene (ATP7A). Genomics 28, 462–469.
Disteche, CM., Gandy, S.L., Adler, D.A. (1987). Translocation and amplification of an X-chromosome repeat in inbred strains of mice. Nucleic Acids Res. 15, 4393–4401.
George, A.M., Reed, V., Glenister, P., Chelly, J., Turner, Z., Horn, N., Monaco, A.P., Boyd, Y. (1994). Analysis of Mnk, the murine homologue of Menke’; disease, in normal and mottled mice. Genomics 22, 27–35.
Green, M.C. (1989). Catalog of mutant genes and polymorphic loci. In Genetic Strains and Variants of the Laboratory Mouse, M.F. Lyon, A.G. Searle, eds. (Oxford: Oxford University Press), pp. 12–403.
Hooper, M., Hardy, K., Handyside, A., Humter, S., Monk, M. (1987). HPRT-deficient (Lesch-Nyhan) mouse embryos derived from germline colonization by cultured cells. Nature 326, 292–295.
Horn, N., Tonnesen, T., Turner, Z. (1992). Menke’; disease: an X-linked neurological disorder of copper metabolism. Brain Pathol. 2, 351–362.
Kaler, S.G., Gallo, L.K., Proud, V.K., Percy, A.K., Mark, Y., Segal, N.A., Goldstein, D.S., Holmes, C.S., Gahl, W.A. (1994). Occipital horn syndrome and a mild Menke’; phenotype associated with splice site mutations at the MNK locus. Nature Genet. 8, 195–202.
Levinson, B., Vulpe, C, Elder, B., Martin, C, Verley, F., Packman, S., Gitschier, J. (1994). The mottled gene is the mouse homologue of the Menkes disease gene. Nature Genet. 6, 369–373.
Mercer, J.F.B., Livingston, J., Hall, B., Paynter, J.A., Begy, C, Chandrasekharappa, S., Lockhart, P., Grimes, A., Bhave, M., Siemieniak, D., Glover, T.W. (1993). Isolation of a partial candidate gene for Menke’; disease by positional cloning. Nature Genet. 3, 20–25.
Mercer, J.F.B., Grimes, A., Ambrosini, L., Lockhart, P., Paynter, J.A., Dierick, H., Glover, T.W. (1994). Mutations in the murine homologue of the Menkes gene in dappled and blotchy mice. Nature Genet. 6, 374–378.
Phillips, R.J.S. (1961). ‘Dappled’, a new allele at the mottled locus in the house mouse. Genet. Res. 2, 290–295.
Rasberry, C, Cattanach, B.M. (1993). Three new mottled mutations (Research News). Mouse Genome 91, 851–853.
Reed, V., Boyd, Y. (1994). Mutations in mottled dappled are RFLVs. Nature Genet. 8, 11–12.
Turner, Z., Vural, B., Tonnesen, T., Chelly, J., Monaco, A.P., Horn, N. (1995). Characterization of the exon structure of the Menke’; disease gene using vectorette PCR. Genomics 26, 437–442.
Vulpe, C, Levinson, B., Whitney, S., Packman, S., Gitschier, J. (1993). Isolation of a candidate gene for Menke’; disease and evidence that it encodes a copper-transporting ATPase. Nature Genet. 3, 7–13.
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Masson, W., Holt, S., Reed, V. et al. The use of compound heterozygotes and Hprt selection to analyze X-linked mottled alleles associated with prenatal lethality. Mammalian Genome 7, 486–489 (1996). https://doi.org/10.1007/s003359900148
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DOI: https://doi.org/10.1007/s003359900148