Barsh G S, Epstein C J
Department of Pediatrics, University of California, San Francisco 94143.
Genomics. 1989 Jul;5(1):9-18. doi: 10.1016/0888-7543(89)90080-3.
Isolation of a gene based on its location, which depends on aligning physical landmarks with the genetic map, can yield basic information about genome structure and organization. As a first step toward isolating the mouse agouti (A) locus, we have begun to define the physical position of this gene relative to genetically linked DNA probes from the Psp, Emv-15, and Src loci. Using a combination of pulsed-field gel techniques that include partial digestion with rare-cutting restriction enzymes and analysis of polymorphic sites present in certain inbred strains, we have constructed long-range restriction maps for each of the probes that span a total of more than 3000 kb. The Src and Emv-15 probes are less than 600 kb apart, but are separated from the Psp probe by at least 1500 kb. By determining the position of a 75-kb deletion that inactivates agouti function, we have localized the A locus to within 500 kb of the Psp probe, but more than 600 kb away from the Emv-15 probe. These physical distances contrast with the known recombination frequencies, 3 +/- 3 cM for A-Psp and less than 0.3 cM for A-Emv-15, and suggest that recombination between A and Emv-15 may be suppressed.
基于基因位置进行基因分离,这依赖于将物理标记与遗传图谱进行比对,能够产生有关基因组结构和组织的基础信息。作为分离小鼠刺鼠(A)基因座的第一步,我们已开始确定该基因相对于来自Psp、Emv - 15和Src基因座的遗传连锁DNA探针的物理位置。使用包括用稀有切割限制酶进行部分消化以及分析某些近交系中存在的多态性位点在内的脉冲场凝胶技术组合,我们为每个探针构建了跨度总计超过3000 kb的长程限制图谱。Src和Emv - 15探针相距不到600 kb,但与Psp探针相隔至少1500 kb。通过确定一个使刺鼠功能失活的75 kb缺失的位置,我们已将A基因座定位在距Psp探针500 kb范围内,但距Emv - 15探针超过600 kb。这些物理距离与已知的重组频率形成对比,A - Psp为3±3 cM,A - Emv - 15小于0.3 cM,这表明A和Emv - 15之间的重组可能受到抑制。