Klockars T, Savukoski M, Isosomppi J, Laan M, Järvelä I, Petrukhin K, Palotie A, Peltonen L
Department of Human Molecular Genetics, National Public Health Institute, Helsinki, 00300, Finland.
Genomics. 1996 Jul 1;35(1):71-8. doi: 10.1006/geno.1996.0324.
The variant form of late infantile neuronal ceroid lipofuscinosis (vLINCL, locus definition CLN5) represents a progressive brain disease with autosomal recessive inheritance. We have previously assigned the CLN5 locus to chromosome 13q21.1-q32 between markers D13S160 and D13S162 by linkage analysis in Finnish families. The information on ancient recombination events obtained from linkage disequilibrium provided an efficient tool for further refining the assignment of the CLN5 locus. Isolation of two novel (CA)n markers, COLAC1 and AC224, resulted in a dramatic restriction of the critical DNA region. We utilized the Fiber-FISH technique to orient and order the large DNA clones isolated by STSs and were able to eliminate almost totally the restriction digestion and PFGE step in the construction of the long-range DNA contig. Both linkage disequilibrium data and Fiber-FISH analyses assigned the CLN5 locus to a well-defined 200-kb region. Here we report a complete physical map of about 350 kb covering the critical chromosomal region of CLN5, which will facilitate the final isolation of the CLN5 gene.
晚发性婴儿神经元蜡样脂褐质沉积症的变异型（vLINCL，基因座定义为CLN5）是一种具有常染色体隐性遗传的进行性脑部疾病。我们之前通过对芬兰家族的连锁分析，将CLN5基因座定位于13号染色体q21.1 - q32区域，介于标记D13S160和D13S162之间。从连锁不平衡获得的古代重组事件信息为进一步精确CLN5基因座的定位提供了有效工具。两个新的（CA）n标记COLAC1和AC224的分离，极大地缩小了关键DNA区域。我们利用纤维荧光原位杂交（Fiber-FISH）技术对通过序列标签位点（STS）分离的大型DNA克隆进行定向和排序，并且在构建长程DNA重叠群时几乎完全消除了限制性酶切和脉冲场凝胶电泳（PFGE）步骤。连锁不平衡数据和纤维荧光原位杂交分析都将CLN5基因座定位于一个明确的200 kb区域。在此我们报告了一个约350 kb的完整物理图谱，覆盖CLN5关键染色体区域，这将有助于最终分离出CLN5基因。
