Generation and analysis of transgenic mice carrying parvoviral NS gene

细小病毒具有小的单链DNA，其非结构蛋白(NS)调节病毒衣壳蛋白(VP)和宿主基因的表达。细小病毒与自身免疫性疾病的相关性或因免疫活性改变而怀疑的抗肿瘤发生。因此，我们尝试用细小病毒NS蛋白修饰自身免疫性疾病和抗肿瘤的转基因小鼠。首先，针对NS对小鼠胚胎的潜在细胞毒性，应用四环素诱导基因表达系统产生过量表达细小病毒NS的转基因小鼠。获得了4只携带反向四环素反式激活因子(RTTA)基因的小鼠和3只携带四环素应答元件(Tre)和细小病毒NS基因的融合转基因小鼠。另一方面，我们还检测了NS的细胞毒性以及细胞因子与NS在宿主细胞中的相互作用，以推测NS过表达的表型。结果表明，NS的细胞毒性与诱导细胞凋亡有关，并且NS与CBP(CREB结合蛋白)相互作用，CBP是结合多种转录因子的重要共激活因子。

Parvoviruses have small single stranded DNA,its nonstructural (NS) protein regulates viral capsid protein (VP) and host gene expressions. Correlation between parvoviruses and autoimmun diseases or antitumorigenesis suspected because of modification of immunocompetence. Thus, we tried to generate transgenic mice to involve modification of autoimmune disease and anti tumorigenesis with parvoviral NS protein.At first, tetracycline inducible gene expression system was applied to generate transgenic mice overexperssing prvoviral NS,because of potential cytotoxicity of NS to mouse fetus. We generated 4 founder mice carrying reverse tetracycline transactivator (rtTA) gene and 3 founder mice carrying fusion transgene including tetracycline responsive element (TRE) and parvoviral NS genes. These mice are cross breeding to generate dual transgenic mice overexpression NS by administration of tetracycline.On the other hand, we also examined cytotoxicity of NS and cellular factors interact with NS in host cells, to expect phenotypes by overexpression of NS.The results indicate that cytotoxicity of NS is correlated with induction of apoptosis, and that NS interacts to CBP (CREB binding protein) which is an important coactivators to bind several transcriptional factors.