Skip to main content
SpringerLink
Log in

Cloning of genomic sequences from the human Y chromosome after purification by dual beam flow sorting

  • Original Investigations
  • Published:
Human Genetics Aims and scope Submit manuscript

Summary

Human Y chromosomes were purified by dual beam flow sorting from a human x Chinese hamster cell line retaining the Y as the only free human chromosome. DNA was extracted from the Y fraction and cloned into λgtWES.λB vector arms. More than 100 recombinant clones carrying human inserts have been characterised by Benton-Davis plaque screening and Southern blotting or in situ hybridisation. Several repetitive sequences were found to be predominantly located on the Y, whereas the majority also cross-hybridised with autosomal DNA. One repetitive clone gave a specific hybridisation signal with the X and the Y chromosome but not with autosomes. Preliminary evidence indicates that many clones contain single copy as well as repetitive sequences. However, no Y-specific single copy sequence has yet been identified.

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

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  • Benton WD, Davis RW (1977) Screening γgt recombinant clones by hybridisation to single plaques in situ. Science 196:180–182

    Google Scholar 

  • Bostock CJ, Gosden JR, Mitchell AR (1978) Localisation of a male specific DNA fragment to a sub-region of the human Y chromosome. Nature 272:324–328

    Google Scholar 

  • Bühler EM (1980) A synopsis of the human Y chromosome. Hum Genet 55:145–175

    Google Scholar 

  • Cooke H (1976) Repeated sequences specific to human males. Nature 262:182–186

    Google Scholar 

  • Cooke HJ, McKay RDG (1978) Evolution of a human Y chromosome-specific repeated sequence. Cell 13:453–460

    Google Scholar 

  • Cremer C, Gray JW, Rappold G, Müller CR, Ropers HH (1983) Preparative dual beam sorting of the human Y chromosome and in situ-hybridisation of cloned DNA probes. Cytometry (in press)

  • Cremer C, Gray JW, Ropers HH (1982) Flow cytometric characterisation of a Chinese hamster x man hybrid cell line retaining the human Y chromosome. Hum Genet 60:262–266

    Google Scholar 

  • Davies KE, Young BD, Elles RG, Hill ME, Williamson R (1980) Cloning of a representative genomic library of the human X chromosome after sorting by flow cytometry. Nature 293:374–376

    Google Scholar 

  • Dean PN (1980) Dual beam sorting at Livermore. In: Laerum OD, Lindmo T, Thorud E (eds) Flow cytometry IV. Universitetsforlaget, Bergen Oslo Tromso, pp 41–44

    Google Scholar 

  • Dean PN, Pinkel D (1978) High resolution dual laser flow cytometry. J Histochem Cytochem 26:622–627

    Google Scholar 

  • Denhardt DT (1966) A membrane-filter technique for the detection of complementary DNA. Biochem Biophys Res Commun 23:641–646

    Google Scholar 

  • Enquist LW, Madden MJ, Schiop-Stansly P, Vande Woude GF (1979) Cloning of Herpes simplex type 1 DNA fragments in a bacteriophage lambda vector. Science 203:541–544

    Google Scholar 

  • Gerhard PS, Kawasaki ES, Bancroft FC, Szabo P (1981) Localisation of a unique gene by direct hybridisation in situ. Proc Natl Acad Sci USA 78:3755–3760

    Google Scholar 

  • Goodfellow P, Banting G, Sheer D, Ropers HH, Caine A, Ferguson-Smith MA, Povey S, Voss R (1983) Genetic evidence that a Y-linked gene in man is homologous to a gene on the X chromosome. Nature 302:346–349

    Google Scholar 

  • Hohn B (1979) In vitro packaging of λ and cosmid DNA. Methods Enzymol 68:299–309

    Google Scholar 

  • Horan PK, Wheeless LL Jr (1977) Quantitative single cell analysis and sorting. Science 198:149–157

    Google Scholar 

  • Kinross J, Fraccaro M, Scapaticci S, Tiepolo L, Zuffardi O, Pawlowitzki IH, Jones KW (1978) Bsu restriction of DNA from cases exhibiting sex chromosome abnormalities. Cytogenet Cell Genet 20:59–69

    Google Scholar 

  • Krumlauf R, Jeanpierre M, Young BD (1982) Construction and characterization of genomic libraries from specific human chromosomes. Proc Natl Acad Sci USA 79:2971–2974

    Google Scholar 

  • Kunkel LM, Smith KD, Boyer SH (1976) Human Y specific reiterated DNA Science 191:1189–1190

    Google Scholar 

  • Kunkel LM, Smith KD, Boyer SH (1979) Organization and heterogeneity of sequences within a repeating unit of human Y chromosome desoxyribonucleic acid. Biochemistry 18:3343–3353

    Google Scholar 

  • Kunkel LM, Smith KD, Boyer SH, Borgaonkar DS, Wachtel SS, Miller OJ, Breg WR, Jones HW, Rary JM (1977) Analysis of human Y-chromosome specific reiterated DNA in chromosome variants. Proc Natl Acad Sci USA 74:1245–1249

    Google Scholar 

  • Kunkel LM, Tantravahi U, Eisenhard M, Latt S (1982) Regional localisation on the human X chromosome of DNA segments cloned from flow sorted chromosomes. Nucleic Acid Res 10: 1557–1578

    Google Scholar 

  • Lawn RM, Fritsch EF, Parker RC, Blake G, Maniatis T (1978) The isolation and characterisation of linked β- and δ-globin genes from a cloned library of human DNA Cell 15:1157–1174

    Google Scholar 

  • Maniatis T, Fritsch EF, Sambrook J (1982) In: Molecular cloning. Cold Spring Harbor Laboratory. Cold Spring Harbor, NY, pp 371–373

    Google Scholar 

  • McKay RDG, Bobrow M, Cooke HJ (1978) The identification of a repeated DNA sequence involved in the karyotype polymorphisms of the human Y chromosome. Cytogenet Cell Genet 21:19–32

    Google Scholar 

  • McKusick VA (1982) The human gene map. Clin Genet 22:359–391

    Google Scholar 

  • Melamed MR, Mullaney PF, Mendelsohn ML (eds) (1979) Flow cytometry and sorting. John Wiley & Sons, New York Chicester Brisbane Toronto

    Google Scholar 

  • Migeon BR, Shapiro LJ, Norum RA, Mohandas T, Axelman J, Dabora RL (1982) Differential expression of steroid sulphatase locus on active and inactive human X chromosome. Nature 299: 838–840

    Google Scholar 

  • Mohandas T, Shapiro LJ, Sparkes RS, Sparkes MC (1979) Regional assignment of the steroid sulfatase-X-linked ichthyosis locus: implications for a non-inactivated region of the short arm of human X chromosome. Proc Natl Acad Sci USA 76:5779–5783

    Google Scholar 

  • Müller CR, Migl B, Traupe H, Ropers HH (1980) X-linked steroid sulfatase: evidence for different gene-dosage in males and females. Hum Genet 54:197–204

    Google Scholar 

  • Page D, de Martinville B, Barker D, Wyman A, White R, Francke U, Botstein D (1982) Single copy sequence hybridises to polymorphic and homologous loci on human X and Y chromosome. Proc Natl Acad Sci USA 79:5352–5356

    Google Scholar 

  • Pearson PL, Bobrow M (1970) Definitive evidence for the short arm of the Y chromosome associating with the X chromosome during meiosis in the human male. Nature 226:959–961

    Google Scholar 

  • Polani P (1982) Pairing of X and Y chromosomes, non-inactivation of X-linked genes, and the maleness factor. Hum Genet 60:207–211

    Google Scholar 

  • Race RR, Sanger R (1975) In: Blood groups in man. Blackwell Scientific Publications. Oxford, pp 578–591

    Google Scholar 

  • Rigby PWJ, Dieckmann M, Rhodes C, Berg P (1976) Labeling deoxyribonucleic acid to high specific activity in vitro by nick translation with DNA polymerase I. J Mol Biol 113:237–251

    Google Scholar 

  • Ropers HH, Wiberg U (1982) Evidence for X-linkage and non-inactivation of steroid sulphatase locus in wood lemming. Nature 296:766–767

    Google Scholar 

  • Schmid CW, Deininger PL (1975) Sequence organisation of the human genome. Cell 6:345–358

    Google Scholar 

  • Solari AJ, Tres LL (1970) The three-dimensional reconstruction of the XY chromosomal pair in human spermatocytes. J Cell Biol 45: 43–53

    Google Scholar 

  • Solari AJ (1980) Synaptonemal complexes and associated structures in microspread human spermatocytes. Chromosoma 81:315–337

    Google Scholar 

  • Southern EMJ (1975) Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol 98:503–511

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Institut für Humangenetik und Anthropologie der Universität, Albertstr. 11, D-7800, Freiburg i. Br., Federal Republic of Germany

    C. R. Müller, C. Cremer, G. Rappold & H. H. Ropers

  2. Biochemistry Department, St. Mary's Hospital Medical School, W2 1PG, London, England

    K. E. Davies

  3. Institut für Anthropologie und Humangenetik der Universität, D-6900, Heidelberg, Federal Republic of Germany

    G. Rappold

  4. Biomedical Science Division, Lawrence Livermore National Laboratory, 94550, Livermore, CA, USA

    J. W. Gray

Authors
  1. C. R. Müller
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. K. E. Davies
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. C. Cremer
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. G. Rappold
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. J. W. Gray
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. H. H. Ropers
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Müller, C.R., Davies, K.E., Cremer, C. et al. Cloning of genomic sequences from the human Y chromosome after purification by dual beam flow sorting. Hum Genet 64, 110–115 (1983). https://doi.org/10.1007/BF00327104

Download citation

  • Received:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00327104

Keywords

Search

Navigation