Springer Nature is making SARS-CoV-2 and COVID-19 research free. View research | View latest news | Sign up for updates

Cytologic and molecular analysis of 46,XXq- cells to identify a DNA segment that might serve as a probe for a putative human X chromosome inactivation center

  • 35 Accesses

  • 41 Citations


Cloned human X chromosome-specific DNA segments, derived from a recombinant phage library enriched for the human X and previously localized to different regions of the X, were used as probes in Southern blots to confirm the nature of a deletion of the long arm of the X chromosome as del (X)(q13) in a patient with some features of Turner's syndrome and suspected from cytologic studies to have a 46,XXq- karyotype. Two dimensional scanning densitometry of autoradiograms of the Southern blots was used to quantitate hybridization of the 32P-labeled probes, reinforcing visual analysis and permitting distinction between sequences present at one or two copies per diploid genome. Once thus characterized, DNA from the patient's cells was used in quantitatively analyzed Southern blots to refine the location of an additional DNA segment, previously mapped to somewhere in the proximal part of the long arm of the X chromosome, to the juxtacentromeric region of Xq, which has been hypothesized to be critical for X-inactivation. Cloned DNA probes such as that localized to the juxtacentromeric region of Xq should be useful for evaluating this hypothesis.

This is a preview of subscription content, log in to check access.


  1. Barr ML, Bertram EG (1949) A morphological distinction between neurons of the male and female, and the behavior of the nucleoton satellite during accelerated nucleoprotein synthesis. Nature 163: 676–677

  2. Biemont MC, Laurent C, Couturier J, Dutrillaux B (1978) Chronologie de la replication des bandes des chromosomes sexuels dans les lymphocytes de sujets normaux et anormaux. Ann Genet (Paris) 21:133–141

  3. Bocian M, Krmpotic E, Szego K, Rosenthal IM (1971) Somatic stigmata of Turner's syndrome in a patient with 46,XXq-. J Med Genet 8: 358–363

  4. Boczkowski K, Mikkelsen M (1973) Fluorescence and autoradiographic studies in patients with Turner's syndrome and 46,XXq- and 46,XXq-karyotypes. J Med Genet 10:350–355

  5. Brunk CF, Jones KC, James TW (1979) Assay for nanogram quantities of DNA in cellular homogenates. Anal Biochem 92:497–500

  6. Bruns GAP, Gusella JF, Keys C, Housman D, Gerald PS (1981) Isolation of DNA segments from the human X chromosome. Pediatr Res 15:599 (Abstract 704)

  7. Czaker R (1973) Banding patterns and late replication in Hela cells. Hum Genet 19:135–144

  8. Daly RF, Patau K, Therman E, Sarto GE (1977) Structure and Barr body formation of an Xp+ chromosome with two inactivation centers. Am J Hum Genet 29:83–93

  9. Davies KE, Young BD, Elles RG, Hill ME, Williamson R (1981) Cloning of a representative genomic library of the human X chromosome after sorting by flow cytometry. Nature 293:374–376

  10. Disteche CM, Eicher EM, Latt SA (1979) Late replication in an X-autosome translocation in the mouse: Correlation with genetic inactivation and evidence for selective effects during embryogenesis. Proc Natl Acad Sci USA 76:5234–5238

  11. Eicher EM (1970) X-autosome translocation in the mouse: Total inactivation versus partial inactivation of the X chromosome. Adv Genet 15:175–259

  12. Epstein CJ, Travis B, Tucker G, Smith S (1978) The direct demonstration of an X-chromosome dosage effect prior to inactivation. In: Russell LB (ed) Genetic mosaics and chimeras in mammals. Plenum Press, New York, pp 261–267

  13. Gartler SM, Andina RJ (1976) Mammalian X-chromosome inactivation. Adv Hum Genet 7:99–140

  14. German JL (1962) DNA synthesis in human chromosomes. Ann NY Acad Sci 24:395–407

  15. Goldman B, Polani PE, Daker MG, Angell RR (1982) Clinical and cytogenetic aspects of X-chromosome deletions. Clin Genet 21: 36–52

  16. Kahan B, DeMars R (1975) Localized derepression on the human inactive X chromosome in mouse-human cell hybrids. Proc Natl Acad Sci USA 72:1510–1514

  17. Kunkel LM, Tantravahi U, Eisenhard M, Latt SA (1982) Regional localization on the human X of DNA segments cloned from flow sorted chromosomes. Nucleic Acids Res 10:1557–1578

  18. Latt SA (1974) Microfluorometric analysis of DNA replication in a human X chromosome. Exp Cell Res 86:412–415

  19. Latt SA (1975) Fluorescence analysis of late DNA replication in human metaphase chromosomes. Somatic Cell Genet 1:293–321

  20. Latt SA (1979) Patterns of late replication in human X chromosomes. In: Vallet HL, Porter IH (eds) Genetic mechanisms of sexual development. Academic Press, New York, pp 305–326

  21. Latt SA, Willard HF, Gerald PS (1976) BrdU-33258 Hoechst analysis of DNA replication in human lymphocytes with supernumerary or structurally abnormal X chromosomes. Chromosoma 57:135–153

  22. Lawn RM, Fritsch EF, Parker RC, Blake G, Maniatis T (1978) The isolation and characterization of linked and B globin genes from a cloned library of human DNA. Cell 15:1157–1174

  23. Lester SC, Korn NJ, Demars R (1982) Derepression of genes on the human inactive X chromosome: Evidence for differences in locus-specific rates of derepression and rates of transfer of active and inactive genes after DNA-mediated transformation. Somatic Cell Genet 8:265–284

  24. Lyon MF (1961) Gene action in the X chromosome of the mouse. Nature 190:372–373

  25. Lyon MF (1971) Possible mechanisms of X-chromosome inactivation. Nature 232:229–232

  26. McCaw BK, Latt SA (1977) X-chromosome replication in parthenogenetic benign ovarian teratomas. Hum Genet 38:253–264

  27. Mattei MG, Mattei JF, Vidal I, Giraud F (1981) Structural anomalies of the X chromosome and inactivation conten. Hum Genet 56: 401–408

  28. Mohandas T, Shapiro LJ, Sparkes RS, Sparkes MS (1979) Regional assignment of the steroid sulfatase-X-linked ichthyosis locus: Implications for a noninactivated region on the short arm of human X chromosome. Proc Natl Acad Sci USA 76:5779–5783

  29. Mohandas T, Sparkes RS, Shapiro LJ (1981) Reactivation of an inactive human X chromosome: Evidence for X inactivation by DNA methylation. Science 211:393–396

  30. O'Farrel P (1981) Replacement synthesis method of labelling DNA fragments. Focus 3:1–3

  31. Rigby PW, Dieckman M, Rhodes C, Berg P (1977) Labeling deoxyribonucleic acid to high specific activity in vitro by nick translation with DNA polymerase. J Mol Biol 113:237–251

  32. Sarto GE, Therman E, Patau K (1974) Increased Q fluorescence of an inactive Xq-chromosome in man. Clin Genet 6:289–293

  33. Seabright M (1971) A rapid banding technique for human chromosomes. Lancet ii:971–972

  34. Shapiro LJ, Mohandas T, Weiss R, Tomeo G (1979) Non-inactivation of an X-chromosome locus in man. Science 204:1224–1226

  35. Slayter EM (1970) Photographic methods in optical instrumentation. In: Optical methods in biology. Wiley-Interscience, New York, pp 460–479

  36. Southern EM (1975) Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol 98:503–517

  37. Sumner AT (1972) A simple technique for demonstrating centromeric heterochromatin. Exp Cell Res 75:304–306

  38. Tantravahi U, Guntaka RV, Erlanger BF, Miller OJ (1981) Amplified ribosomal RNA genes in a rat hepatoma cell line are enriched in 5-methylcytosine. Proc Natl Acad Sci USA 78:489–493

  39. Therman E, Sarto GE, Patau K (1974) Center for Barr body condensation on the proximal part of the human Xq: A hypothesis. Chromosoma 44:361–366

  40. Therman E, Sarto GE, Palmer CG, Kallio H, Denniston C (1979) Position of the human X inactivation center on Xq. Hum Genet 50:59–64

  41. Therman E, Denniston C, Sarto GE, Ulber M (1980) X chromosome constitution and the human female phenotype. Hum Genet 54: 133–134

  42. Westway D, Williamson D (1981) An intron nucleotide sequence variant in a cloned B+ thalassemia globin gene. Nucleic Acids Res 9: 1777–1788

  43. Willard HF (1977) Tissue-specific heterogeneity in DNA replication in human X chromosomes. Chromosoma 61:61–73

  44. Willard HF, Latt SA (1976) Analysis of deoxyribonucleic acid replication in human X chromosomes by fluorescence microscopy. Am J Hum Genet 28:213–227

  45. Wilard HF, Smith KD, Goss SJ (1981) Identification and regional localization of an X-specific reiterated DNA fragment from the human X chromosome. Am J Hum Genet 33:59A

  46. Wolf SF, Mareni CE, Migeon BR (1980) Isolation and characterization of cloned DNA sequences that hybridize to the human X chromosome. Cell 21:95–102

  47. Wyss D, DeLozier CD, Daniell J, Engel E (1982) Structural anomalies of the X chromosome: Personal observation and review of nonmosaic cases. Clin Genet 21:145–159

Download references

Author information

Correspondence to U. Tantravahi.

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Tantravahi, U., Kirschner, D.A., Beauregard, L. et al. Cytologic and molecular analysis of 46,XXq- cells to identify a DNA segment that might serve as a probe for a putative human X chromosome inactivation center. Hum Genet 64, 33–38 (1983). https://doi.org/10.1007/BF00289475

Download citation


  • Southern Blot
  • Analyze Southern Blot
  • Visual Analysis
  • Proximal Part
  • Cytologic Study