Lerner T J, D'Arigo K L, Haines J L, Doggett N A, Taschner P E, de Vos N, Buckler A J
Molecular Neurogenetics Unit, Massachusetts General Hospital, Charlestown 02129, USA.
Am J Med Genet. 1995 Jun 5;57(2):320-3. doi: 10.1002/ajmg.1320570243.
In order to identify genes originating from the Batten disease candidate region, we have used the technique of exon amplification to identify transcribed sequences. This procedure produces trapped exon clones, which can represent single exons or multiple exons spliced together and is an efficient method for obtaining probes for physical mapping and for screening cDNA libraries. The source of DNA for these experiments was a collection of chromosome 16 cosmid contigs isolated by the direct subcloning of region-specific yeast artificial chromosomes (YACs) and hybridization of inter-alu PCR products from these YACs to the flow-sorted Los Alamos chromosome 16 cosmid library. We are now using the resulting exon probes to screen retina and brain cDNA libraries for candidate JNCL genes.
为了鉴定源自巴顿病候选区域的基因,我们采用外显子扩增技术来鉴定转录序列。该程序产生捕获的外显子克隆,其可以代表单个外显子或拼接在一起的多个外显子,并且是获得用于物理图谱绘制和筛选cDNA文库的探针的有效方法。这些实验的DNA来源是通过区域特异性酵母人工染色体(YAC)的直接亚克隆以及来自这些YAC的间α-PCR产物与经流式细胞术分选的洛斯阿拉莫斯16号染色体粘粒文库杂交而分离得到的16号染色体粘粒重叠群集合。我们现在正在使用所得的外显子探针筛选视网膜和脑cDNA文库以寻找候选的JNCL基因。