Blanco G, Nikitopoulou A, Kraus M, Mason R M, Coulton G R, Brown S D
MRC Mammalian Genetics Unit and UK Mouse Genome Centre, Harwell, Oxon, OX11 7US, United Kingdom.
Genomics. 1998 Dec 15;54(3):415-23. doi: 10.1006/geno.1998.5614.
The ky mouse mutant exhibits a degenerative muscle disease resulting in chronic deformation of the spinal column. Following a previous report describing the mapping of the ky locus to a small region of mouse chromosome 9 (Skynner et al., 1995, Genomics 25, 207-213), we have now undertaken a positional cloning approach to identify candidate genes for ky. A YAC/BAC contig encompassing the ky locus was constructed comprising 48 YAC clones and 48 newly generated STSs. The results from the combined physical and genetic analyses showed that only two overlapping BAC clones, which together do not exceed 260 kb, span the ky nonrecombinant region. A combination of gene hunting methods on the critical BACs has led to the identification of seven coding fragments, which have been tested for expression. The expression analysis and the position of the coding fragments on the contig suggest their grouping in at least four transcription units. One of these transcription units is expressed exclusively in skeletal muscle, making it a suitable candidate for this muscle defect in the ky mouse.
ky小鼠突变体表现出一种退行性肌肉疾病，导致脊柱慢性变形。继之前一份将ky基因座定位到小鼠9号染色体一个小区域的报告（Skynner等人，1995年，《基因组学》25卷，207 - 213页）之后，我们现在采用了定位克隆方法来鉴定ky的候选基因。构建了一个包含ky基因座的YAC/BAC重叠群，其中包括48个YAC克隆和48个新生成的STS。物理和遗传分析的结果表明，只有两个重叠的BAC克隆（其总长不超过260 kb）跨越了ky非重组区域。对关键BAC进行的一系列基因搜寻方法已鉴定出七个编码片段，并对其表达进行了检测。表达分析以及编码片段在重叠群上的位置表明它们至少可分为四个转录单位。其中一个转录单位仅在骨骼肌中表达，这使其成为ky小鼠这种肌肉缺陷的合适候选基因。
