Development of new SNP typing technologies for searching disease-associated genes.

开发用于搜索疾病相关基因的新 SNP 分型技术。

• 批准号: 12557237
• 负责人: TOKUNAGA Katsushi
• 金额: $ 7.04万
• 依托单位: The University of Tokyo
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 2000
• 资助国家: 日本
• 起止时间: 2000 至 2002
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-12557237/
• 关键词: single nucleotide polymorphism (SNP); SNP typing technology; DNA micro array; disease associated gene; rheumatoid arthritis; systemic lupus erythematosus; DNAマイクロキャピラリーアレイ; 一分子蛍光検出法; HLA遺伝子; SNP; DNAアレイ

In order to establish reliable technologies for typing nucleotide sequence polymorphisms, we first developed a new typing system for the HLA-DRB1 gene, which shows a highest degree of polymorphism and significant associations with many diseases. A glass array with a chamber made the typing procedure similar to that for microtiter plates. Next we developed a micro-capillary array for the typing of disease-associated SNPs. Many different oligonucleotide probes were spotted in each capillary, and each probe provided a specific hybridization signal after washing through a capillary. Thus this system may be suitable for multiplex and automated SNP typing. In addition, we could establish a new SNP typing system in the combination of PCR-SSP with the fluorescence correlation spectroscopy. The system has been introduced to routine SNP typing and proved to be a large-scale and cost-effective system. Furthermore, we have identified several rheumatic disease-associated SNPs including those of FCGR2B, FCGR3A and LIR1 genes.

为了建立可靠的核苷酸序列多态性分型技术，我们首先开发了一种新的HLA-DRB 1基因分型系统，该基因显示出最高程度的多态性，并与许多疾病有显着关联。带有腔室的玻璃阵列使分型过程类似于微量滴定板。接下来，我们开发了一种用于疾病相关SNP分型的微毛细管阵列。在每个毛细管中点样许多不同的寡核苷酸探针，并且每个探针在通过毛细管洗涤后提供特异性杂交信号。因此，该系统可适用于多重和自动化SNP分型。此外，PCR-SSP与荧光相关光谱技术相结合，可以建立一种新的SNP分型系统。该系统已被引入常规SNP分型，并被证明是一个大规模和成本效益的系统。此外，我们已经确定了一些风湿性疾病相关的SNP，包括FCGR 2B，FCGR 3A和LIR 1基因。

项目成果

Kato H, et al.: "Single nucleotide polymorphisms in the coding region of human CXC-chemokine receptors CXCR1, CXCR2 and CXCR3"Genes immun.. 1. 330-337 (2000)

Kato H 等：“人 CXC 趋化因子受体 CXCR1、CXCR2 和 CXCR3 编码区的单核苷酸多态性”Genes immun.. 1. 330-337 (2000)

Tsuchiya N, Kawasaki A, Tsao BP, Komata T, Grossman JM, Tokunaga K: "Analysis of the association of HLA-DRBl, TNFα promoter and TNFR2 (TNFRSF1B) polymorphisms with SLE using transmission disequilibrium test"Genes Immun.. 2・6. 317-322 (2001)

Tsuchiya N、Kawasaki A、Tsao BP、Komata T、Grossman JM、Tokunaga K：“使用传递不平衡测试分析 HLA-DRB1、TNFα 启动子和 TNFR2 (TNFRSF1B) 多态性与 SLE 的关联”Genes Immun.. 2・6 .317-322 (2001)

Shibue T,Tsuchiya N,Komata T, et al.: "Tumor necrosis factor α 5'-flanking region, TNF receptor II, and HLA-DRB1 polymorphisms in the Japanese patients with rheumatoid arthritis."Arthritis Rheum.. 43・3. 753-757 (2000)

Shibue T、Tsuchiya N、Komata T 等人：“日本类风湿关节炎患者的肿瘤坏死因子 α 5 侧翼区、TNF 受体 II 和 HLA-DRB1 多态性。”关节炎大黄.. 43・3。 753-757 (2000)

Kawasaki A, et al.: "Independent contribution of HLA-DRB1 and TNFa promoter polymorphisms to the susceptibility to Crohn^ユs disease"Genes Immun.. 1. 351-357 (2000)

Kawasaki A 等人：“HLA-DRB1 和 TNFa 启动子多态性对克罗恩病易感性的独立贡献” Genes Immun.. 1. 351-357 (2000)

Kawasaki A, et al.: "Analysis on the association of human BLYS (BAFF, TNFSF13B)polymorphisms with systemic lupus erythematosus and rheumatoid arthritis"Genes. Immun.. 3(7). 424-429 (2002)

Kawasaki A 等人：“人类 BLYS（BAFF、TNFSF13B）多态性与系统性红斑狼疮和类风湿性关节炎的关联分析”基因。