Kunz J, Scherer S W, Klawitz I, Soder S, Du Y Z, Speich N, Kalff-Suske M, Heng H H, Tsui L C, Grzeschik K H
Institut für Humangenetik, Philipps-Universität Marburg, Germany.
Genomics. 1994 Jul 15;22(2):439-48. doi: 10.1006/geno.1994.1407.
Toward the construction of a complete physical map of human chromosome 7, we have localized 725 YAC clones to cytogenetically defined regions using fluorescence in situ hybridization (FISH) and by screening with DNA markers of known chromosomal locations. These chromosome 7-specific YAC clones are part of a library constructed with DNA isolated from monochromosomal 7 human-hamster somatic cell hybrid lines. The FISH mapping for 575 clones was accomplished by using "Alu-PCR" amplified YAC DNA against metaphase chromosome spreads made from a monochromosomal 7 human-mouse somatic cell hybrid line. Hybridization- or PCR-based screening of previously mapped DNA markers was performed for the mapping of 221 YAC clones. There was excellent correlation between the map locations obtained for the 71 YACs localized with both methods. All of the regionally localized YAC clones are valuable reagents for mapping and identification of disease genes on human chromosome 7.
为构建人类7号染色体的完整物理图谱,我们利用荧光原位杂交(FISH)并通过用已知染色体位置的DNA标记进行筛选,将725个酵母人工染色体(YAC)克隆定位到细胞遗传学定义的区域。这些7号染色体特异性YAC克隆是用从单条7号染色体的人-仓鼠体细胞杂种系分离的DNA构建的文库的一部分。575个克隆的FISH定位是通过使用“Alu-PCR”扩增的YAC DNA与由单条7号染色体的人-小鼠体细胞杂种系制备的中期染色体铺片杂交来完成的。对221个YAC克隆的定位则通过基于杂交或PCR的先前定位的DNA标记筛选来进行。用两种方法定位的71个YAC所获得的图谱位置之间存在极好的相关性。所有区域定位的YAC克隆都是用于人类7号染色体疾病基因定位和鉴定的有价值试剂。