Deidda G, Cacurri S, Piazzo N, Felicetti L
Istituto di Biologia Cellulare, CNR, Rome, Italy.
J Med Genet. 1996 May;33(5):361-5. doi: 10.1136/jmg.33.5.361.
The p13E-11 probe has been shown to detect DNA rearrangements in sporadic and familial cases of FSHD. Its use, however, has been hampered by the fact that it detects at least two pairs of EcoRI alleles, one derived from the 4q35 region (D4F104S1), the other from 10q26 (D10F104S2). We have cloned p13E-11 EcoRI fragments from the 4q35 and 10q26 subtelomeric regions and shown the presence of several restriction site differences within the KpnI tandem repeat units. The two loci present a different distribution of restriction sites for the enzyme BlnI which allows differential cleavage of the KpnI units derived from 10q26, leaving intact the 4q35 pair of alleles. This method of differential restriction greatly facilitates the interpretation of Southern blots obtained from affected and unaffected subjects, with an important improvement in reliability for diagnosis and genetic counselling. In addition, this method can be used to investigate the molecular mechanism of the 4q35 rearrangement implicated in the disease and to ascertain whether the rearrangement is because of interchromosomal exchange between 4qter and 10qter KpnI repeats.
p13E - 11探针已被证明可检测散发性和家族性面肩肱型肌营养不良（FSHD）病例中的DNA重排。然而，它的应用受到了限制，因为它能检测至少两对EcoRI等位基因，一对来自4q35区域（D4F104S1），另一对来自10q26（D10F104S2）。我们从4q35和10q26亚端粒区域克隆了p13E - 11 EcoRI片段，并显示在KpnI串联重复单元内存在几个限制性酶切位点差异。这两个位点对BlnI酶呈现不同的限制性酶切位点分布，这使得源自10q26的KpnI单元能够被特异性切割，而4q35等位基因对保持完整。这种差异限制性酶切方法极大地促进了对患病和未患病个体Southern印迹结果的解读，在诊断和遗传咨询的可靠性方面有了重要提高。此外，该方法可用于研究与该疾病相关的4q35重排的分子机制，并确定这种重排是否是由于4q末端和10q末端KpnI重复序列之间的染色体间交换所致。
