Kurimasa A, Suzuki N, Kumano S, Li H, Wells D, Wagner M J, Chen F, Chen D J, Oshimura M
Department of Molecular and Cell Genetics, School of Life Science, Faculty of Medicine, Tottori University, Japan.
Genomics. 1995 Jul 20;28(2):147-53. doi: 10.1006/geno.1995.1125.
Microcell hybrids containing various regions of human chromosome 8 were formed by microcell-mediated transfer of neo-tagged chromosome 8 into the cells derived from severe combined immunodeficiency (SCID) mouse. Thus, 110 cosmid markers were isolated from SV40-transformed SCID fibroblast cell line (SCVA) containing a p12-q11.1 region of human chromosome 8 and were assigned to eight regions in 8p12-q11.1, using a microcell-hybrid panel. For positional cloning of a human gene that restores the DNA-repair defect in a mouse with SCID on 8p11.1-q11.1 (SCID region), we constructed a yeast artificial chromosome (YAC) contig of about 4.5 Mb. Overlapping YACs were further aligned by restriction mapping, using rare-cutting restriction endonucleases. The cosmids and YAC contig should facilitate isolation of the SCID gene and other genes, such as the Werner syndrome-responsible gene in or near this region.
通过微细胞介导将新标记的8号染色体转移到严重联合免疫缺陷（SCID）小鼠来源的细胞中，形成了包含人类8号染色体不同区域的微细胞杂种。因此，从含有人类8号染色体p12-q11.1区域的SV40转化的SCID成纤维细胞系（SCVA）中分离出110个黏粒标记，并利用微细胞杂种细胞板将其定位到8p12-q11.1的八个区域。为了对位于8p11.1-q11.1（SCID区域）上能恢复SCID小鼠DNA修复缺陷的人类基因进行定位克隆，我们构建了一个约4.5 Mb的酵母人工染色体（YAC）重叠群。使用稀有切割限制内切酶通过限制酶切图谱对重叠的YAC进行进一步比对。这些黏粒和YAC重叠群将有助于分离SCID基因以及该区域内或附近的其他基因，如沃纳综合征相关基因。
