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Isolation of anonymous DNA markers for human chromosome 22q11 from a flow-sorted library, and mapping using hybrids from patients with DiGeorge syndrome

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Summary

DiGeorge syndrome (DGS) is a human developmental defect of the structures derived from the third and fourth pharyngeal pouches. It apparently arises due to deletion of 22q11. We describe a strategy for the isolation of DNA probes for this region. A deleted chromosome 22, which includes 22q11, was flow-sorted from a lymphoblastoid cell line of a patient with cat eye syndrome and used as the source of DNA. A DNA library was constructed from this chromosome by cloning into the EcoR1 site of the vector Lambda gt10. Inserts were amplified by PCR and mapped using a somatic cell hybrid panel of this region. Out of 32 probes, 14 were mapped to 22q11. These probes were further sublocalised within the region by dosage analysis of DGS patients, and by the use of two new hybrid cell lines which we have produced from DGS patients. One of these lines (7939B662) contains the altered human chromosome segregated from its normal homologue. This chromosome 22 contains an interstitial deletion in 22q11, and will be useful for localising further probes to the DGS region.

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References

  • Buckton KE, Spowart G, Newton MS, Evans HJ (1985) Fortyfour probands with an additional marker chromosome. Hum Genet 69:353–370

    Google Scholar 

  • Carey AH, Roach S, Williamson R, Dumanski JP, Nordensjold M, Collins VP, Rouleau G, Blin N, Jalbert P, Scambler PJ (1990) Localisation of 27 DNA markers to the region of chromosome 22q11-pter, deleted in patients with the DiGeorge syndrome and duplicated in the der22 syndrome. Genomics 7:299–306

    Google Scholar 

  • Chapelle A de la, Herva R, Koivisto M, Aula P (1981) A deletion in chromosome 22 can cause DiGeorge syndrome. Hum Genet 57:253–256

    Google Scholar 

  • Chisaka O, Capecchi MR (1991) Regionally restricted developmental defects resulting from targeted disruption of the mouse homeobox gene hox-1. 5. Nature 350:473–479

    Google Scholar 

  • Conley ME, Beckwith JB, Mancer JFK, Tenckhoff L (1979) The spectrum of the DiGeorge syndrome. J Pediatr 94:883–890

    Google Scholar 

  • DiGeorge AM (1965) Discussions on a new concept of the cellular basis of immunology. J Pediatr 67:907–908

    Google Scholar 

  • Dumanski JP (1990) Rapid procedures for the isolation of novel chromosome-specific DNA probes from a phage library. Technique 2:31–35

    Google Scholar 

  • Dumanski JP, Geurts van Kessel AHM, Ruttledge M, Wladis A, Sugawa N, Collins PV, Nordenskjold M (1990) Isolation of anonymous polymorphic DNA fragments from human chromosome 22q12-qter. Hum Genet 84:219–222

    Google Scholar 

  • Emmanuel BS (1988) Molecular cytogenetics: towards dissection of contiguous gene syndromes. Am J Hum Genet 43:575–578

    Google Scholar 

  • Faed MJW, Robertson J, Swanson Beck J, Cater JI, Base B, Madlorn M (1987) Features of DiGeorge syndrome in a child with 45,XX-3,-22,+der(3),t(3;22)(p25;q1. 1). J Med Genet 24:225–234

    Google Scholar 

  • Fames JA, Green DK, Malloy P, Sumner AT (1989) Flow cytometry measurement of kinetochore labelling. Cytometry 10:134–142

    Google Scholar 

  • Feinberg AP, Vogelstein B (1983) A technique for labelling restriction endonuclease fragments to high specific activity. Anal Biochem 132:6–13

    CAS  PubMed  Google Scholar 

  • Fibison WJ, Budarf M, Mcdermid H, Greenberg F, Emanuel BS (1990) Molecular studies of DiGeorge syndrome. Am J Hum Genet 46:888–895

    Google Scholar 

  • Fuscoe JC, Clark LM, Van Dilla MA (1986) Construction of 15 chromosome-specific libraries from flow purified chromosomes. Cytogenet Cell Genet 43:79–86

    Google Scholar 

  • Geurts van Kessel AHM, Westerveld A, DeGroot PG, Meera Khan P, Hagemeijer A (1980) Regional localisation of genes coding for human ACO2, ARSA, and NAGA on chromosome 22. Cytogenet Cell Genet 28:169–172

    Google Scholar 

  • Greenberg F, Crowder WE, Paschall V, Colon-luares JC, Lubianski B, Ledbetter D (1984) Familial DiGeorge syndrome and associated partial monosomy of chromosome 22. Hum Genet 65:317–319

    Google Scholar 

  • Greenberg F, Valdes C, Rosenblatt HM, Kirkland JL, Ledbetter DH (1986) Hypoparathyroidism and T-cell immune defect in a patient with 10p deletion syndrome. J Pediatr 109:489–492

    Google Scholar 

  • Greenberg F, Elder FFB, Haffner P, Northrup H, Ledbetter DH (1988) Cytogenetic findings in a prospective series of patients with DiGeorge anomaly. Am J Hum Genet 43:605–611

    Google Scholar 

  • Hofker MJ, Breuning MJ, Bakker E, Van Ommen GJB, Pearson PL (1985) An anonymous single-copy chromosome 22 clone D22S10 (22cl-18) identifies an RFLP with Pst1. Nucleic Acids Res 19:7167

    Google Scholar 

  • Jorgensen AL, Kolvraa S, Jones C, Lethbak A (1988) A subfamily of alphoid repetitive DNA shared by the NOR-bearing human chromosomes 14 and 22. Genomics 3:100–109

    Google Scholar 

  • Lammer EJ, Opitz JM (1986) The DiGeorge anomaly as a developmental field defect. Am J Med Genet Suppl 2:113–127

    Google Scholar 

  • Lodewyk H, Van Mierop S, Kutsche LM (1986) Cardiovascular anomalies in DiGeorge syndrome, and importance of neural crest as a possible pathogenic factor. Am J Cardiol 58:133–137

    Article  CAS  PubMed  Google Scholar 

  • Lotscher E, Grzeschik K-H, Bauer HG, Pohlenz H-D, Straubinger B, Zachau HG (1986) Dispersed human Immunoglobulin kappa light chain genes. Nature 320:456–458

    Google Scholar 

  • Mascarello JT, Bastian JF, Jones MC (1989) Interstitial deletion of chromosome 22 in a patient with DiGeorge malformation sequence. Am J Med Genet 32:112–114

    Google Scholar 

  • McDermid HE, Duncan AMV, Brash KR, Holden JJA, Magenis E, Sheehy R, Burn J, Karden N, Noel B, Schinzel A, Teshima I, White BN (1986) Characterisation of the supernumary chromosome in cat eye syndrome. Science 232:646–648

    Google Scholar 

  • Metzdorf R, Gottert E, Blin N (1988) A novel centromeric repetitive DNA from human chromosome 22. Chromosoma 97:154–158

    Google Scholar 

  • Mitchell AR, Gosden JR, Miller DA (1985) p82H: a cloned sequence of the alphoid repeated family found at the centromeres of all human chromosomes. Chromosoma 92:369–377

    Google Scholar 

  • Pinto M, Leite RP, Areias A (1989) Features of Turner and Di-George syndromes in a child with an X:22 translocation. J Med Genet 26:778–780

    Google Scholar 

  • Scambler PJ, Carey AH, Wyse RKH, Roach S, Dumanski JP, Nordenskjold M, Williamson R (1991) Microdeletions within 22q11 associated with sporadic and familial DiGeorge syndrome. Genomics 10:201–206

    Google Scholar 

  • Sealey PG, Whittaker PA, Southern EM (1985) Removal of repeated sequences from hybridisation probes. Nucleic Acids Res 13:1905–1922

    Google Scholar 

  • Van Heyningen V, Bickmore WA, Seawright A, Fletcher JM, Maule J, Fekete G, Gessler M, Bruns GA, Huerre-Jeanpiere C, Junien C, Williams BRG, Hastie ND (1990) Role for the Wilms tumour gene in genital development. Proc Natl Acad Sci USA 87:5383–5386

    Google Scholar 

  • Van Keuren ML, Hart IM, Kao FT, Neve RL, Bruns GAP, Kurnit DM, Patterson D (1987) A somatic cell hybrid with a single human chromosome 22 corrects the defect in the CHO mutant (Ade-1) lacking adenylosuccinase activity. Cytogenet Cell Genet 44:142–147

    Google Scholar 

  • Zhang FR, Delaitre O, Rouleau G, Couturier J, Lefrancois D, Thomas G, Aurias A (1990) The neuroepithelioma breakpoint on chromosome 22 is proximal to the meningioma locus. Genomics 6:174–177

    Google Scholar 

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Sharkey, A.M., McLaren, L., Carroll, M. et al. Isolation of anonymous DNA markers for human chromosome 22q11 from a flow-sorted library, and mapping using hybrids from patients with DiGeorge syndrome. Hum Genet 89, 73–78 (1992). https://doi.org/10.1007/BF00207046

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