STRUCTURE AND FUNCTION OF NOVEL GENE (BRG1) EXPRESSED IN PANCREATIC BETA CELLS

胰腺β细胞表达的新基因（BRG1）的结构和功能

基本信息

批准号：
11671131
负责人：
HAMAGUCHI Kazuyuki
金额：
$ 1.92万
依托单位：
OITA MEDICAL UNIVERISTY
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
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

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-11671131/
关键词：
pancreatic β cell differential display type 1 diabetes MIN6 TNF-α 1型糖尿病 HLA スピードコンジェニック NODマウス cDNA

项目摘要

Utilizing the differential display technique with arbitrarily-primed PCR, we obtained a 〜500bp fragment, which is preferentially amplified in the cDNA from mouse β cell lines. We named the gene as BRG1 (β related gene 1). The mRNA was very large, I.e. 〜14kb, and the levels of expression were higher in the groups cultured with high glucose than low glucose in MIN6 cells and rat islets. Tissue distribution of BRG1 mRNA expression revealed that BRG1 gene was expressed ubiquitously in various mouse tissues. The highest expression was observed in the testes. The almost entire region of 〜14kb cDNA was sequenced and determined. There were 87〜89 % sequence identities between the mouse and the human sequences. On the other hand, sequence identities between mouse and Drosophila BRG1 sequence were 67〜74 % in each of 5 restricted regions of about 84〜324 bp stretches. The deduced BRG1 amino acid sequences for mouse and human (〜4, 400 A.A.) had 93.8 % identity throughout the entire region. On the other hand, there was 41.8 % identity in the C-terminal 〜3,800 amino acid region between mouse and Drosophila BRG1 protein. Moreover, there was 26.8 % identity in the C-terminal 〜1,650 amino acid region between mouse and C. elegans BRG1 protein. The chromosomal mapping of the BRG1 gene using the mouse RH panel revealed that the BRG1 is located at the distal part of mouse chromosome 4, where one of the diabetogenic genes in NOD mouse, Iddll, has been mapped.The second research theme was about the polymorphisms in the 5'-flanking region of the tumor necrosis factor (TNF)-α gene in Japanese type 1 diabetes. We observed that the TNFP-D and -B alleles were associated with type 1 diabetes, which may be secondary to their linkage disequilibria with the susceptible HLA class I and class II alleles.

利用差异显示技术和任意引物PCR，我们获得了约500bp的片段，该片段在小鼠β细胞系的cDNA中优先扩增，我们将该基因命名为BRG1（β相关基因1）。即~14kb，并且在MIN6细胞和大鼠胰岛中高糖培养组中的表达水平高于低糖培养组中的BRG1 mRNA表达的组织分布。 BRG1 基因在小鼠的各种组织中普遍表达，在测试中观察到几乎整个区域的 cDNA 与人类序列之间有 87-89% 的序列同一性。另一方面，推断出小鼠和果蝇BRG1序列在约84〜324bp延伸的5个限制区中的每一个中的序列同一性为67〜74%。小鼠和人类的 BRG1 氨基酸序列（约 4, 400 个氨基酸）在整个区域具有 93.8% 的同一性，而小鼠和果蝇 BRG1 蛋白的 C 端约 3,800 个氨基酸区域具有 41.8% 的同一性。此外，小鼠和线虫之间 C 端 ~1,650 个氨基酸区域有 26.8% 的同一性BRG1 蛋白。使用小鼠 RH panel 对 BRG1 基因进行染色体定位，发现 BRG1 位于小鼠 4 号染色体的远端，NOD 小鼠的糖尿病基因之一 Iddll 已被定位于此。第二个研究主题关于日本 1 型糖尿病肿瘤坏死因子 (TNF)-α 基因 5' 侧翼区域的多态性我们观察到 TNFP-D 和-B 等位基因与 1 型糖尿病相关，这可能是由于它们与易感 HLA I 类和 II 类等位基因的连锁不平衡所致。