Functional analysis of HCV core protein using HCV core transgenic mouse

使用HCV核心转基因小鼠进行HCV核心蛋白的功能分析

基本信息

批准号：
11670476
负责人：
IMAZEKI Fumio
金额：
$ 2.3万
依托单位：
Chiba University
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 2000
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-11670476/
关键词：
HCV core protein transgenic mouse suppression subtractive hybridization cDNA array DNA chip mRNA トランスジェニックマウス C型肝炎ウイルスコア蛋白サブトラクション法肝細胞癌 DNAチップ C型慢性肝炎 Northern blotting C型肝炎ウイルス HCVコアタンパク Fasシグナル伝達系細胞周期細胞増殖

项目摘要

We analyzed comprihensively the difference of mRNAs expressed in the livers of HCV core transgenic mouse and wild mouse in order to clarify the function of HCV core protein in vivo by the method of subtraction and cDNA microarray. We have got fifty clones of HCV core transgenic mouse subtracted from wild one and 44 clones of wild mouse subtracted from HCV core transgenic one by the method of suppression subtractive hybridization. Sequence analysis of these clones showed not any genes associated with inflammation or carcinogenesis, and the intensity of expression of these genes between transgenic and wild ones was within two times. Next, we analyzed the difference of expression of about 1000 genes between transgenic and wild mice by cDNA array and DNA chip, respectively. Both methods revealed several genes expressed more than two times in transgenic mouse, and these genes seem also not associated with inflammation or carcinogenesis. We will analyze the significance of these genes from now.Expression of HCV core mRNA was ascertained in the transgenic mouse used in this study by northern blotting and the amount of mRNA was estimated to be around 2 pg/μgRNA in the liver. HCV core protein was not analyzed in the taransgenic mouse used in this study, but previous results showed the expression of HCV core protein by western blotting. Therefore HCV core protein might have little effect on the expression of genes in this transgenic mouse in vivo.

我们全面分析了在HCV核心转基因小鼠和野生小鼠生活中表达的mRNA差异，以通过减法和cDNA微阵列的方法阐明HCV核心蛋白在体内的功能。通过抑制减去杂交方法，我们从野生小鼠中减去了五十个HCV核转基因小鼠的HCV核转基因小鼠，从HCV核心转基因中减去野生小鼠的克隆和44个克隆。这些克隆的序列分析没有显示与感染或癌变有关的任何基因，而这些基因在转基因和野生基因之间的表达强度在两次之内。接下来，我们分别通过cDNA阵列和DNA芯片分析了转基因和野生小鼠之间约1000个基因的表达差。两种方法都揭示了几种基因在转基因小鼠中表达了两次以上，这些基因似乎也与感染或癌变无关。我们将从现在开始分析这些基因的重要性。在本研究中使用的转基因小鼠中，通过北印迹确定了HCV核mRNA的表达，据估计，肝脏中的mRNA量约为2 pg/μgrna。在这项研究中使用的taransgic小鼠中未分析HCV核心蛋白，但先前的结果表明，HCV核心蛋白通过蛋白质印迹的表达。因此，HCV核心蛋白可能对该转基因小鼠在体内的基因表达几乎没有影响。