Hori S, Sewry C A, Dubowitz V, Morris G E
MRIC Biotechnology Group, N.E. Wales Institute, Deeside, Clwyd, United Kingdom.
Am J Med Genet. 1995 Aug 28;58(2):177-86. doi: 10.1002/ajmg.1320580217.
We have produced a new panel of 20 monoclonal antibodies (mAbs) against a region of the dystrophin protein corresponding to a deletion-prone region of the Duchenne muscular dystrophy gene (exons 45-50). We show that immuno-histochemistry or Western blotting with these "exon-specific" mAbs can provide a valuable addition to Southern blotting or PCR methods for the accurate identification of genetic deletions in Becker muscular dystrophy patients. The antibodies were mapped to the following exons: exon 45 (2 mAbs), exon 46 (6), exon 47 (1), exons 47/48 (4), exons 48-50 (6), and exon 50 (1). PCR amplification of single exons or groups of exons was used both to produce specific dystrophin immunogens and to map the mAbs obtained. PCR-mediated mutagenesis was also used to identify regions of dystrophin important for mAb binding. Because the mAbs can be used to characterize the dystrophin produced by individual muscle fibres, they will also be useful for studying "revertant" fibres in Duchenne muscle and for monitoring the results of myoblast therapy trials in MD patients with deletions in this region of the dystrophin gene.
我们制备了一组新的20种单克隆抗体（mAb），它们针对肌营养不良蛋白的一个区域，该区域对应于杜兴氏肌营养不良基因的一个易于缺失的区域（外显子45 - 50）。我们表明，用这些“外显子特异性”单克隆抗体进行免疫组织化学或蛋白质印迹分析，可为Southern印迹法或PCR方法提供有价值的补充，用于准确鉴定贝克肌营养不良患者的基因缺失。这些抗体被定位到以下外显子：外显子45（2种单克隆抗体）、外显子46（6种）、外显子47（1种）、外显子47/48（4种）、外显子48 - 50（6种）和外显子50（1种）。单个外显子或外显子组的PCR扩增既用于产生特异性肌营养不良蛋白免疫原，也用于定位获得的单克隆抗体。PCR介导的诱变也用于鉴定肌营养不良蛋白中对单克隆抗体结合重要的区域。由于这些单克隆抗体可用于表征单个肌纤维产生的肌营养不良蛋白，它们也将有助于研究杜兴氏肌中的“回复”纤维，并监测肌营养不良蛋白基因该区域存在缺失的MD患者的成肌细胞治疗试验结果。
