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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.

12Q染色体上DARIER病基因区物理图谱的构建并尝试检测患者DNA上该区域的基因组缺失。

基本信息

批准号：
08670983
负责人：
IKEDA Shigaku
金额：
$ 1.6万
依托单位：
JUNTENDO UNIVERSITY,SCHOOL OF MEDICINE
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
1996
资助国家：
日本
起止时间：
1996 至 1997
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-08670983/
关键词：
DARIER DISEASE POSITIONAL CLONING GENE PHYSICAL MAP

项目摘要

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.

1993年，通过对欧洲和中东祖先的遗传连锁分析，将达里尔病(DAR)的基因定位于ch12q23-24.1。从那时起，几位研究人员做出了巨大的努力来克隆该基因。因此，在1996-2007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)我们对患有这种疾病的日本家庭进行了测试，发现在日本家庭中，座位异质性也可能不常见。(2)通过检测几个新分离的多态DNA标记，将DAR亚组分成D12S1328和D12S1616之间的2兆碱基区间。(3)建立了包含DAR区域的基因组重叠群。重叠群的着丝粒和端粒大部分由我们自己建立，中心部分由我们的共生代谢家建立(Pulst S.et al：Natural Genetics，14：269,1996)。(3)对丝氨酸/苏氨酸蛋白磷酸酶基因的外显子-内含子连接进行了测序，并对部分患者进行了SSCP分析。到目前为止，还没有发现异常情况。我们现在正在测试另外几名患者的运动检测。(4)为了分离候选基因，我们对重叠群上的一些基因组克隆进行了外显子捕捉法。在初步程序中检测到了一些表达序列，目前正在进行进一步的测试，以确定可能的候选范围。(5)最后，用从重叠群获得的几个探针，对26例无关患者的基因组DNA样本进行了脉冲场电泳+Southern杂交，以鉴定基因组缺失。到目前为止，还没有发现异常情况。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。