Identification of the genes for human diseases by the nucleotide analyses in wide range and developing the model mouse for them

通过广泛的核苷酸分析鉴定人类疾病的基因并为其开发模型小鼠

基本信息

批准号：
11470125
负责人：
KIMURA Minoru
金额：
$ 5.44万
依托单位：
Tokai University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
1999
资助国家：
日本
起止时间：
1999 至 2001
项目状态：
已结题

项目摘要

Based on our previous achievement that HLA region (spanning 1.8 Mb) positioned at the short arm of 6p21. 3 in the human chromosome 6 has already been sequenced with high accuracy, we attempted to identify candidate genes responsible for two human diseases (Bechet's disease [BA] and Psoriasis-[PA]) and create animal models for these two diseases. The following is the achievement we have performed in the past three years.1. A region responsible for PA was confined to a region spanning 89 to 143 kb from the telomere side of HLA-C locus by genetic analyses. In this region, there were one known and 4 novel genes expressed in a skin. Sequencing of this region revealed that one of the 4 novel gene exhibited a significant polymorphism. This gene was termed "SEEK1" and found to overexpress in the skin suffered with PA.2. For the candidate gene for BD, HLA-B gene itself or its neiboring MICA or MICB gene has been considered for the best candidate. However, recent genetic analysis revealed that HLA-B51 gene is the responsible gene for BD.3. We produced several transgenic (Tg) lines carrying a 30-kb fragment containing SEEK1 gene. Further analysis of these lines is now underway.4. We have already produced Tg lines carrying MICA or MICB gene, just prior to decision that HLA-B51 gene is the responsible gene for BD. These MICA or MICB Tg lines exhibited several abnormalities such as reduction in the nubmer of leukocytes and body weight, and hyperkeratosis in the skin, suggesting possible involvement of these genes in the pathogenesis of BD.5. We also produced Tg lines carrying HLA-B51 and human b2-microglobulin genes and found that in these mice expression of HLA-B51 molecules on the cell surface was greatly elevated. Mice carrying human b2-microglobulin (b2m) and HLA-B51 genes, but lacking mouse b2m gene are now successfully produced using b2m-knock out mice.

基于我们以前的成就，即HLA地区（跨越1.8 MB）位于6p21的短臂。 3在人类染色体上已经以很高的精度进行了测序，我们试图识别造成两种人类疾病（Bechet病[BA]和牛皮癣 - [PA]）的候选基因，并为这两种疾病创建动物模型。以下是我们在过去三年中取得的成就1。通过遗传分析，将负责PA的区域局限于HLA-C基因座端粒侧89至143 kb的区域。在该地区，有一个在皮肤中表达的已知和4个新基因。该区域的测序表明，四个新型基因之一表现出显着的多态性。该基因被称为“ Seek1”，发现在皮肤中过表达Pa.2。对于BD的候选基因，HLA-B基因本身或其Neiboring Mica或MICB基因已被视为最佳候选者。然而，最近的遗传分析表明，HLA-B51基因是BD.3的负责基因。我们产生了几种含有30 kb片段的转基因（TG）线，其中包含SEEK1基因。现在正在进行对这些线路的进一步分析4。在决定HLA-B51基因是BD的负责基因之前，我们已经产生了带有云母或MICB基因的TG系。这些云母或MICB TG系表现出多种异常，例如减少白细胞和体重的努伯，以及皮肤中的高乳变性，表明这些基因可能参与BD.5的发病机理。我们还产生了带有HLA-B51和人B2-微球蛋白基因的TG系，发现在这些小鼠中，在细胞表面上HLA-B51分子的表达大大升高。携带人B2-微球蛋白（B2M）和HLA-B51基因但缺乏小鼠B2M基因的小鼠现在使用B2M敲除小鼠成功产生。