Mammalian Genome

, Volume 5, Issue 11, pp 663–669 | Cite as

Deleted Chromosome 20 from a patient with Alagille syndrome isolated in a cell hybrid through leucine transport selection: study of three candidate genes

  • J. F. Deleuze
  • S. Dhorne
  • J. Hazan
  • E. Borghi
  • N. Raynaud
  • N. Pollet
  • M. Meunier-Rotival
  • J. Deschatrette
  • D. Alagille
  • M. Hadchouel
Original Contributions


Alagille syndrome (AGS) is a well-defined genetic entity assigned to the short arm of Chromosome (Chr) 20 by a series of observations of AGS patients associated with microdeletions in this region. By fusing lymphoblastoid cells of an AGS patient that exhibited a microdeletion in the short arm of Chr 20 encompassing bands p11.23 to p12.3 with rodent thermosensitive mutant cells (CHOtsH1-l) deficient in-leucyl-tRNA synthetase, we isolated a somatic cell hybrid segregating the deleted human Chr 20. This hybrid clone, designated NR2, was characterized by several methods, including PCR, with eight pairs of oligonucleotides mapped to Chr 20: D20S5, D20S41, D20S42, D20S56, D20S57, D20S58, adenosine deaminase (ADA), and Prion protein (PRIP); Restriction Fragment Length Polymorphism (RFLP) analyses with four genomic anonymous probes (D20S5, cD3H12, D20S17, D20S18); and fluorescent in situ hybridization (FISH) with total human DNA and D20Z1, a sequence specific to the human Chr 20 centromere, as probes.

The NR2 hybrid allowed us to exclude three candidate genes for AGS: hepatic nuclear factor 3 β (HNF3β), paired box 1 (PAX1), and cystatin C (CST3) as shown by their localization outside of the deletion. The NR2 hybrid is a powerful tool for the mapping of new probes of this region, as well as for obtaining new informative probes specific for the deletion by subtractive cloning of the region. Such markers will be useful for linkage analysis and screening of cDNA libraries.


Prion Protein Cell Hybrid Adenosine Deaminase Somatic Cell Hybrid Hybrid Clone 
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Copyright information

© Springer-Verlag New York Inc 1994

Authors and Affiliations

  • J. F. Deleuze
    • 1
  • S. Dhorne
    • 1
  • J. Hazan
    • 2
  • E. Borghi
    • 3
  • N. Raynaud
    • 1
  • N. Pollet
    • 1
  • M. Meunier-Rotival
    • 1
  • J. Deschatrette
    • 1
  • D. Alagille
    • 1
  • M. Hadchouel
    • 1
  1. 1.INSERM U 347 et Département de PédiatrieLe Kremlin Bicêtre CedexFrance
  2. 2.Unité de génétique moléculaire humaineCNRS URA 1445, Institut PasteurParis Cedex 15France
  3. 3.Laboratoire de biologie de la reproduction et du développementHôpital de Bicêtrele Kremlin Bicêtre CedexFrance

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