腺病毒介导的人巨细胞病毒UL49基因小鼠模型的建立

建立表达HCMV UL49 基因的转基因小鼠，为抗病毒药物研究提供有效的实验动物模型。本实验将UL49-GFP基因插入腺病毒穿梭质粒pDC316中,构建重组质粒pDC316-UL49-GFP，与腺病毒骨架质粒pBHGloxΔE1,3Cre 通过脂质体介导共转染293 细胞，重组产生腺病毒Ad-UL49-GFP, 经PCR和Western Blot鉴定正确后，大量扩增、纯化，制备高滴度重组腺病毒。纯化腺病毒经尾静脉注射感染小鼠，通过荧光定量PCR 和Western blot 方法，检测UL49 基因在小鼠体内组织分布和表达时相。结果显示UL49基因在小鼠的心、肝、脾、肺、肾组织均有表达，并且表达量由高到低顺序依次是：肝、脾、肾、心、肺，在腺病毒感染第3天在各靶器官表达水平较高，此后逐渐下降，第14天时仅存在肝和脾中。表明表达UL49基因的小鼠模型构建成功。小鼠模型的成功建立为下一步筛选以UL49基因为靶的抗病毒药物奠定了基础。

Recombinant Adenovirus Vector Mediated Expression of HCMV UL49 Gene in Mice

Human cytomegalovirus (HCMV) is extremely species specific and does not replicate in experimental animal tissues.To overcome the problem and establish suitable animal models for studying antiviral strategies,the expression of HCMV UL49 gene was explored in mice.UL49-GFP gene was subcloned into the adenovirus shuttle plasmid pDC316,the products(pDC316-UL49-GFP)were co-transfected with helper plasmid pBHGloxE1,3Cre into HEK293 cell lines by liposome reagent,recombinant adenovirus(Ad-UL49-GFP) was generated an...