To generate new chromosome 21 markers in a region that is critical for the pathogenesis of Down syndrome (D21S55-MX1), we used pulsed field gel electrophoresis (PFGE) to isolate a 600-kb NruI DNA fragment from the WA17 hybrid cell line, which has retained chromosome 21 as the only human material. This fragment, which contains the oncogene ETS2, was used to construct a partial genomic library. Among the 14 unique sequences that were isolated, 3 were polymorphic markers and contained sequences that are conserved in mammals. Five of these markers mapped on the ETS2-containing NruI fragment and allowed us to define an 800-kb high-resolution PFGE map.
This is a preview of subscription content, log in to check access.
Buy single article
Instant access to the full article PDF.
Price includes VAT for USA
Subscribe to journal
Immediate online access to all issues from 2019. Subscription will auto renew annually.
This is the net price. Taxes to be calculated in checkout.
Albertsen HM, LePaslier H, Abderrahim J, Dausset J, Cann H, Cohen D (1989) Improved control of partial DNA restriction enzyme digest in agarose using limiting concentrations of Mg++. Nucleic Acids Res 17:808
Anand R, Honeykombe J, Whittaker PA, Elder JK, Southern EM (1988) Clones from an 850Kb fragment containing the 5′ region of the DMD locus enriched by pulsed field gel electrophoresis. Genomics 3:177–186
Carle CF, Olson MV (1984) Separation of chromosomal DNA molecules from yeast by orthogonal-field-alteration gel electrophoresis. Nucleic Acids Res 12:5647–5664
Créau-Goldberg N, Delabar JM, Crété N, Sinet PM (1989) Partial physical map of human chromosome 21. Cytogenet Cell Génét 51:981–982
Crété N, Delabar JM, Sinet PM, Créau-Goldberg N (1991a) Accurate evaluation of the sizes of DNA fragments (from 30 to 4700 Kb) in pulsed field gel electrophoresis. BioTechniques 11:712–718
Crété N, Delabar J, Yaspo ML, Sinet PM, Créau-Goldberg N (1991b) Five groups of physical linkage on chromosome 21. Cytogenet Cell Genet 58:2035
Deininger PL, Jolly DL, Rubin CM, Friedman T, Schmid CW (1981) Base sequence studies of 300 nucleotide renatured repeated human DNA clones. J Mol Biol 151:17–33
Gardiner K (1990) Pulsed field gel electrophoresis and investigations into mammalian genome organization. J Cell Sci 96:5–8
Gardiner K, Patterson D (1988) Transverse alternating electrophoresis. Nature 331:371–372
Gardiner K, Patterson D (1989) Transverse alternating field electrophoresis and applications to mammalian genome mapping. Electrophoresis J 10:296–302
Gardiner K, Laas W, Patterson D (1986) Fractionation of large mammalian DNA restriction fragments using vertical pulsed field gradient gel electrophoresis. Somatic Cell Mol Genet 12:185–195
Gardiner K, Horisberger M, Kraus J, Tantravahi U, Korenberg J, Rao V, Reddy S, Patterson D (1990) Analysis of human chromosome 21:correlation of physical and cytogenetic maps; gene and CpG island distributions. EMBO J 9:25–34
Michiels F, Burmeister M, Lehrach H (1987) Derivation of clones close to met by preparative field inversion gel electrophoresis. Science 236:1305–1308
Rahmani Z, Blouin JL, Créau-Goldberg N, Watkins P, Mattei JF, Poissonnier M, Prieur M, Chettouh Z, Nicole A, Aurias A, Sinet PM, Delabar J (1989) Critical role of the D21S55 region on chromosome 21 in the pathogenesis of Down syndrome. Proc Natl Acad Sci USA 86:5958–5962
Ramsay M, Wainwright BJ, Farral M, Estivill X, Sutherland H, Ho MF, Davies R, Halford S, Tata F, Wicking C, Lench N, Bauer I, Ferec C, Farndon P, Kruyer H, Stanier P, Williamson R, Scambler PJ (1990) A new polymorphic locus, D7S411, isolated by cloning from preparative pulsa-field gels is close to the mutation causing cystic fibrosis. Genomics 6:39–47
Raziuddin Arati, Sarkar FH, Dutkowski R, Shulman L, Ruddle FH, Gupta SL (1984) Receptors for human alpha and beta interferon but not gamma interferon are specified by human chromosome 21. Proc Natl Acad Sci USA 81:5504–5508
Schwartz DC, Cantor (1984) Separation of yeast chromosomesized DNAs by pulsed field gel electrophoresis. Cell 37:67–75
Sealey PG, Whittaker PA, Southern EM (1985) Removal of repeated sequences from hybridization probes. Nucleic Acids Res 13:1905–1922
Second international workshop on human chromosome 21 (1991) Cytogenet Cell Genet 57:167–174
Sinet PM, Rahmani Z, Théophile D, Chettouh Z, Blouin JL, Prieur M, Noel B, Pangalos C, Mattei JF, Kraus J, Delabar J (1991) Molecular definition of 7 minimal regions on chromosome 21 involved in the pathogenesis of 23 features of Down syndrome. Cytogenet Cell Genet 58:2040
VanKeuren ML, Watkins PC, Drabkin HA, Jabs EW, Gusella JF, Patterson D (1986) Regional localisation of DNA sequences on chromosome 21 using somatic cell hybrids. Am J Hum Genet 38:793–804
Wong C, Kazazian HH, Stetten G, Earnshaw WC, VanKeuren ML, Antonarakis SE (1989) Molecular mechanism in the formation of human ring chromosome 21. Proc Natl Acad Sci USA 86:1914–1918
About this article
Cite this article
Yaspo, M., Crété, N., Chettouh, Z. et al. New chromosome 21 DNA markers isolated by pulsed field gel electrophoresis from an ETS2-containing Down syndrome chromosomal region. Hum Genet 90, 427–434 (1992). https://doi.org/10.1007/BF00220472
- Internal Medicine
- Metabolic Disease
- Down Syndrome
- Chromosomal Region
- Polymorphic Marker