Functional analysis of HCV core protein using HCV core transgenic mouse

为了进一步阐明HCV核心蛋白在体内的功能，我们采用差减法和基因芯片技术，对HCV核心蛋白转基因小鼠和野生小鼠肝脏中mRNA表达的差异进行了深入分析。用抑制消减杂交法从野生型小鼠中获得了50个HCV核心转基因小鼠克隆，从HCV核心转基因小鼠中获得了44个野生型小鼠克隆。对这些克隆的序列分析表明，没有任何与炎症或致癌相关的基因，这些基因的表达强度在转基因和野生型之间的两倍之内。其次，我们分别用cDNA阵列和DNA芯片分析了转基因小鼠与野生小鼠约1000个基因的表达差异。这两种方法都揭示了一些基因在转基因小鼠中表达超过两倍，并且这些基因似乎也与炎症或致癌无关。用北方印迹法（Northern blotting）检测了转基因小鼠肝脏中HCV核心区mRNA的表达，其表达量约为2 pg/μgRNA。在本研究中使用的taransgenic小鼠中未分析HCV核心蛋白，但先前的结果显示通过蛋白质印迹法表达HCV核心蛋白。因此，HCV核心蛋白可能对该转基因小鼠体内基因表达的影响不大。

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.