Murakami T, Lupski J R
Department of Molecular and Human Genetics, Baylor College of Medicine, One Baylor Plaza, Houston, Texas, 77030, USA.
Genomics. 1996 May 15;34(1):128-33. doi: 10.1006/geno.1996.0251.
Charcot-Marie-Tooth disease type 1A (CMT1A) is associated with a 1. 5-Mb tandem duplication in chromosome 17p11.2-p12, and hereditary neuropathy with liability to pressure palsies (HNPP) is associated with a 1.5-Mb deletion at this locus. Both diseases appear to result from an altered copy number of the peripheral myelin protein-22 gene, PMP22, which maps within the critical region. To identify additional genes and characterize chromosomal elements, a 1.5-Mb cosmid contig of the CMT1A duplication/HNPP deletion critical region was assembled using a yeast artificial chromosome (YAC)-based isolation and binning strategy. Whole YAC probes were used for screening a high-density arrayed chromosome 17-specific cosmid library. Selected cosmids were spotted on dot blots and assigned to bins defined by YACs. This binning of cosmids facilitated the subsequent fingerprint analysis. The 1.5-Mb region was covered by 137 cosmids with a minimum overlap set of 52 cosmids assigned to 17 bins and 9 contigs.
1A型夏科-马里-图斯病（CMT1A）与17号染色体p11.2 - p12区域的1.5兆碱基串联重复相关，而遗传性压力易感性神经病（HNPP）与该位点的1.5兆碱基缺失相关。这两种疾病似乎都是由于外周髓磷脂蛋白22基因（PMP22）的拷贝数改变所致，该基因定位于关键区域内。为了鉴定其他基因并表征染色体元件，使用基于酵母人工染色体（YAC）的分离和分箱策略构建了CMT1A重复/HNPP缺失关键区域的1.5兆碱基黏粒重叠群。使用完整的YAC探针筛选高密度排列的17号染色体特异性黏粒文库。将选定的黏粒点样于点杂交膜上，并分配到由YAC定义的箱中。这种黏粒分箱便于后续的指纹分析。1.5兆碱基区域由137个黏粒覆盖，最少有52个黏粒的重叠集被分配到17个箱和9个重叠群中。
