Den Dunnen J T, Grootscholten P M, Bakker E, Blonden L A, Ginjaar H B, Wapenaar M C, van Paassen H M, van Broeckhoven C, Pearson P L, van Ommen G J
Department of Human Genetics, State University Leiden, The Netherlands.
Am J Hum Genet. 1989 Dec;45(6):835-47.
We have studied 34 Becker and 160 Duchenne muscular dystrophy (DMD) patients with the dystrophin cDNA, using conventional blots and FIGE analysis. One hundred twenty-eight mutations (65%) were found, 115 deletions and 13 duplications, of which 106 deletions and 11 duplications could be precisely mapped in relation to both the mRNA and the major and minor mutation hot spots. Junction fragments, ideal markers for carrier detection, were found in 23 (17%) of the 128 cases. We identified eight new cDNA RFLPs within the DMD gene. With the use of cDNA probes we have completed the long-range map of the DMD gene, by the identification of a 680-kb SfiI fragment containing the gene's 3' end. The size of the DMD gene is now determined to be about 2.3 million basepairs. The combination of cDNA hybridizations with long-range analysis of deletion and duplication patients yields a global picture of the exon spacing within the dystrophin gene. The gene shows a large variability of intron size, ranging from only a few kilobases to 160-180 kb for the P20 intron.
我们使用传统印迹法和FIGE分析，对34例贝克型肌营养不良症患者和160例杜氏肌营养不良症（DMD）患者进行了肌营养不良蛋白cDNA研究。共发现128个突变（65%），其中115个缺失和13个重复，其中106个缺失和11个重复可相对于mRNA以及主要和次要突变热点精确定位。连接片段是携带者检测的理想标志物，在128例病例中的23例（17%）中被发现。我们在DMD基因内鉴定出8个新的cDNA RFLP。通过使用cDNA探针，我们通过鉴定一个包含该基因3'端的680 kb SfiI片段，完成了DMD基因的长程图谱。现在确定DMD基因的大小约为230万个碱基对。cDNA杂交与缺失和重复患者的长程分析相结合，得出了肌营养不良蛋白基因中外显子间距的整体情况。该基因的内含子大小差异很大，从仅几千碱基到P20内含子的160 - 180 kb不等。
