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Functional analisis of invgene which determines lef-right axis and relates to renal development

决定左右轴并与肾脏发育相关的inv基因的功能分析

基本信息

批准号：
12671054
负责人：
TSUCHIYA Ken
金额：
$ 2.43万
依托单位：
Tokyo Woman's Medical University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
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

项目摘要

Vertebrate organisms have a common left-right asymmetry of their visceral organs. However, all unpaired organs of the chest and abdomen, such as heart, stomach, spleen and liver, develop from the midline in the fetus and localize to their normal positions in the adult. The mirror image reversal of this asymmetry is called situs inversus. The inversion of embryonic turning (tnv) mutation in a mouse was created by random insertional mutagenesis. The phenotype of the inv mouse is a consistent mirror-image reversal of the left-right polarity (situs inversus) and cystic formation of the kidneys. In previous project, we succeeded in cloning the whole sequence of mouse inv. As protein encoded by a candidate gene involved 15 consecutive repeats of ankyrin motif, there may be a possibility that a cytoskeletal abnormality was involved in the mechanism and production of structural abnormalities, inversion and cyst formation in the kidney. Based on the sequencing data, we had new results as to inv gene summarized below :1) We succeeded in cloning the human inv gene and determined genomic organization, chromosomal mapping.2) We modified mouse inv gene with some reporter genes, such as GFP protein or lacZ, and inserted into the expression vectors. We intend to use expression vector mounted whole mouse inv gene for transfection analysis in the culture cells.3) We modified mouse inv gene with some reporter genes, such as GFP protein or lacZ, and excluded exon 1 to 2 for made up targeting vector for knockout mouse.

脊椎动物的内脏器官有一个共同的左右不对称。然而，胸腹部的所有未配对器官，如心、胃、脾和肝，在胎儿时从中线发育，在成人时定位于它们的正常位置。这种不对称性的镜像反转被称为逆位。用随机插入突变法在小鼠中产生了胚胎翻转倒位突变。inv小鼠的表型是肾脏左右极性的一致镜像反转（反转部位）和囊性形成。在上一个项目中，我们成功地克隆了小鼠inv基因的全序列。由于候选基因编码的蛋白质涉及锚蛋白基序的15个连续重复，因此可能存在细胞骨架异常参与肾脏结构异常、倒位和囊肿形成的机制和产生的可能性。根据测序结果，我们对inv基因的研究取得了以下新的成果：1）成功克隆了人inv基因，并确定了基因组结构和染色体定位; 2）用GFP蛋白或lacZ等报告基因修饰小鼠inv基因，并将其插入到表达载体中。3）用GFP蛋白或lacZ等报告基因修饰小鼠inv基因，并剔除其第1 ~ 2外显子，构建基因敲除小鼠的靶向载体。