TARGETED DISRUPTION OF CERULOPLASMIN GENE, AND ITS INFLUENCE ON PANCREATIC B CELL FUNCTION

铜蓝蛋白(Cp)基因被证明与遗传性CP缺乏症(HCD)有关，HCD是一种以神经异常为特征的常染色体隐性遗传病。在许多HCD病例中，2型糖尿病(DM)是该病的首发症状，10-20年后，在40-60岁时出现神经异常。因此，我们假设CP基因是DM的易感基因，尽管目前参与DM发病机制尚不清楚。为了阐明这一假说和机制，如果是真的，我们首先尝试制作一只靶向CP基因中断的小鼠(CP基因敲除小鼠：CP-/-小鼠)。从小鼠基因组文库中分别筛选出2个和4个含有外显子1和外显子2-4的克隆。将这些克隆用于DNA构建。最后，我们构建了敲除CP基因外显子2和3的DNA结构。该DNA构建物含有pGK-neo和nLacZ-polA可供选择。我们将此DNA结构引入ES细胞，制成基因敲除小鼠。

The ceruloplasmin (Cp) gene has been shown to be responsible for hereditary Cp deficiency (HCD), which is an autosomal recessive disease characterized by neurological abnormalities. In many HCD cases, type 2 diabetes (DM) was the first symptom of the disease, and 10 - 20 years later at age 40 - 60 the neurological abnormalities occurred. Therefore, we made hypothesis that the Cp gene is a susceptible gene for DM, although the mechanisms involved in the pathogenesis of the development of DM are not clear at present. To clarify this hypothesis an, if it is true, the mechanisms, we first tried to make a mouse, which has targeted disruption of Cp gene (Cp knock out mouse: Cp -/-mouse). Then, this mouse can be used to clarify the mechanisms involved in the pathogenesis of the development of DM.From a mouse genomic library, we screened 2 and 4 clones containing exon 1 and exons 2-4, respectively. These clones were used to make DNA construct. We finally made a DNA construct to knock out exons 2 and 3 of the Cp gene. This DNA construct contains PGK-neo and nLacZ-polyA for selection. We introduced this DNA construct to ES cells to make knock out mouse.