Sharan S K, Holdener-Kenny B, Ruppert S, Schedl A, Kelsey G, Rinchik E M, Magnuson T
Department of Genetics, Case Western Reserve University, Cleveland, Ohio 44106.
Genetics. 1991 Nov;129(3):825-32. doi: 10.1093/genetics/129.3.825.
Previous complementation analyses with five (c11DSD, c5FR60Hg, c2YPSj, c4FR60Hd, c6H) of the mouse albino deletions defined at least two genes on chromosome 7, known as eed and exed, which are necessary for development of the embryonic and extraembryonic ectoderm, respectively, of early postimplantation embryos. The region of chromosome 7 containing these two genes has now been accessed at the molecular level by cloning two of the deletion breakpoint-fusion fragments. The c2YPSj breakpoints were isolated by cloning an EcoRI fragment containing a copy of an albino region-specific repeat unique to c2YPSj DNA. Similarly, the c11DSD breakpoints were isolated by cloning a c11DSD EcoRI fragment detected by a unique-sequence probe mapping proximal to the albino-coat-color locus. By mapping the cloned breakpoints relative to the remaining three deletions, the c11DSD distal breakpoint was found to define the distal limit of the region containing eed, whereas the c2YPSj and c6H distal breakpoints were found to define the proximal and distal limits, respectively, of the region containing exed.
先前对小鼠白化缺失中的五个(c11DSD、c5FR60Hg、c2YPSj、c4FR60Hd、c6H)进行的互补分析确定了7号染色体上至少两个基因,即eed和exed,它们分别是植入后早期胚胎的胚胎外胚层和胚外外胚层发育所必需的。现在,通过克隆两个缺失断点融合片段,在分子水平上研究了7号染色体上包含这两个基因的区域。通过克隆一个含有c2YPSj DNA特有的白化区域特异性重复序列拷贝的EcoRI片段,分离出了c2YPSj断点。同样,通过克隆一个由位于白化毛色位点近端的单序列探针检测到的c11DSD EcoRI片段,分离出了c11DSD断点。通过将克隆的断点相对于其余三个缺失进行定位,发现c11DSD远端断点确定了包含eed区域的远端界限,而c2YPSj和c6H远端断点分别确定了包含exed区域的近端和远端界限。