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DOI 10.1007/s00439-007-0362-y
ORIGINAL INVESTIGATION
Received: 22 February 2007 / Accepted: 21 March 2007 / Published online: 14 April 2007
Springer-Verlag 2007
Introduction
Abnormal O-linked glycosylation of -dystroglycan is the
common pathogenic mechanism in a group of patients with
a clinical spectrum ranging from severe congenital muscular
Present Address:
P. K. Grewal
Department of Cellular and Molecular Medicine,
University of California, San Diego, LA, USA
Present Address:
D. Beltrn-Valero de Bernab
Department of Physiology and Biophysics,
University of Iowa College of Medicine, Iowa City, IA, USA
Present Address:
C. B. Michielse
Faculty of Science, University of Amsterdam,
Amsterdam, The Netherlands
Present Address:
T. Voit
Institut de Myologie, Groupe Hospitalier Piti-Salptrire,
Paris, France
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The breakpoint-spanning region was ampliWed by longrange PCR ampliWcation with speciWc primers (supplementary Table 3) and LA TaqTM (TaKaRa Bio Inc., Shiga,
Japan) using PCR conditions recommended by the manufacturer. We then used diVerent combinations of primers to
further restrict the breakpoint-spanning region and identiWed the breakpoint by direct sequencing.
By long-range PCR ampliWcation we obtained a deletionspanning fragment of 3.5 kb from patient 2. Sequence
analysis of this fragment revealed the 5 and 3 breakpoints,
and a deletion size of 63.1 kb (Fig. 4). This genomic deletion predicts a 239 bp deletion in the transcript resulting in
a shift of the open reading frame within the Wrst predicted
catalytic domain, coding for 29 alternative codons followed
by a premature stop codon, which most likely results in nonsense-mediated decay of the transcript. Unfortunately no
cell-line or tissue sample was available to test this hypothesis. Only the aVected individual was homozygous for the
deletion, the parents and Wve of the seven analyzed
unaVected sibs were heterozygous carriers of the deletion.
No signiWcant match between the 5 and 3 junction
sequences is present, therefore the LARGE deletion is likely
the result of non-homologous end-joining as reported also
for the Large deletion in myd mice (Browning et al. 2005).
Results
Discussion
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