CONSTRUCTION OF THE PHYSICAL MAP AT DARIER DISEASE GENE REGION ON CHROMOSOME 12Q AND AN ATTEMPT TO DETECT THE GENOMIC DELETION IN THIS REGION ON THE DNA OF PATIENTS.

1993年，通过对欧洲和中东祖先种类的遗传连锁分析，将达里尔病（DAR）基因定位为ch12q23-24.1。从那时起，几位研究人员为克隆该基因做出了巨大的努力。然而，到目前为止，这种基因还没有被克隆出来。因此，在1996- 1997年，我们提供了被认为是克隆DAR所必需的基本数据。我们的数据总结如下：(1)我们对日本家族进行了本病检测，发现基因座异质性在日本家族中可能也不常见。(2)通过检测多个新分离的多态DNA标记，DAR亚定位于D12S1328和D12S1616之间的两个兆碱基区间。(3)建立了包含DAR区域的基因组序列。基因组的着丝粒和端粒部分主要由我们建立，而中心部分由我们的合作者建立（Pulst S.et al .: Nature Genetics, 14: 269,1996）。(3)对丝氨酸/苏氨酸蛋白磷酸酶基因的外显子-内含子连接处进行了测序，部分患者进行了SSCP分析。到目前为止，还没有发现异常。我们正在对另外几名患者进行基因突变检测。(4)为了分离“候选”基因，我们对部分基因组克隆进行了外显子捕获方法。在初步程序中检测到一些表达的序列，目前正在进行进一步的测试，以划定可能的“候选”范围。(5)最后，对26例无亲缘关系患者的基因组DNA样本进行了脉冲场电泳+ Southern杂交检测，并从该序列中获得了多个探针，以确定基因组缺失。到目前为止，还没有发现异常。我们正在分离更多的探针，这些探针将用于检测一些患者DNA中的基因组缺失。

In 1993, the gene for Darier's disease (DAR) was mapped to ch12q23-24.1 by genetic linkage analysis of the kindreds of European and Middie-eastern ancestory. Since then, tremendous effort has been made to clone the gene by several researchers. As yet, however, the gene has not been cloned.Accordingly, in 1996-7, we provided basic data thought to be essential for the cloning of DAR.The summary of our data is as follows ; (1) We have tested Japanese family with this disease, and found that locus heterogeneity might be also uncommon in Japanese kindred. (2) By testing several newly isolated polymorhic DNA markers, DAR sublocalized to two megabase interval between D12S1328 and D12S1616.(3) A genomic contig encompassing the DAR region has been established. The most centromeic and telomeric portions of the contig were mainly established by us, while the central portion was established by our collabolators (Pulst S.et al : Nature Genetics, 14 : 269,1996).(3) The exon-intron junctions of serine/threonine protein phosphatase gene, while appeared to locallze to the DAR region, were sequenced, and SSCP analysis was performed in some patients. As yet, abnormalities have not been found. We are now testing for muation detection in additional several patients. (4) In order to isolate "candidate" genes, we have performed the exon-trapping method on some genomic clones on the contig. A few expressed sequnces were detected in the preliminary procedure, and tests are now underway furthetr to delimit the range of possible 'candidates'.(5) Finally, genomic DNA samples from 26 unrelated patients have been tested to identity genomic deletion using pulse-field electrophoresis + Southern hybridization with several probes obtained from the contig. As yet, abnormalities have not been found. We are in the process of isolating additional probes, which will be used to detect genomic deletion in the DNA of some patiants.