鸭viperin基因的克隆、表达及Viperin蛋白的抗病毒活性

摘要

Viperin is a kind of host cellular protein induced by type I and type II interferon, which is found in a wide range of organisms, and is highly conserved in evolution. In this research, we had cloned a full-length open reading frame ( ORF) of duck viperin gene from peripheral blood mononuclear cells using RT-PCR. The viperin gene was successful-ly subcloned into eukaryotic expression vector pEGFP-N1-viperin and transfected into DF-1 cells. After challenged with duck Tembusu virus, the virus replication was analyzed between pEGFP-N1-viperin transfection group and empty vector transfec-tion group. The results showed that duck viperin gene was amplified and containing 1092 bp. Transfection results showed that eukaryotic expression vector pEGFP-N1-viperin was successfully expressed in DF-1 cells. Viperin was able to limit the infection of duck Tembusu virus and the virus infection was significant lower than that of the control group. These results indicated that the duck Viperin had well antiviral activity, and it would lay foundations for further research for antiviral biological functions of Viperin.%Viperin蛋白是一种由Ⅰ型和Ⅱ型干扰素诱导的宿主蛋白,广泛存在于各类动物体内,在进化上高度保守.本研究以鸭外周血淋巴细胞为原材料,通过RT-PCR扩增得到viperin基因,同时构建真核表达质粒pEGFP-N1-viperin,转染至DF-1细胞,用坦布苏病毒攻击后,检测该病毒在转染质粒组和转染空载体组细胞中的增殖情况.结果表明,鸭viperin基因开放阅读框为1092 bp,编码蛋白由363个氨基酸构成.转染结果显示,viperin基因在DF-1细胞中成功表达.坦布苏病毒攻击后,与转染空载体组相比,转染pEGFP-N1-viperin组显著抑制了坦布苏病毒在DF-1细胞中的增殖.说明鸭Viperin蛋白作为抗病毒蛋白,具有良好的抑制坦布苏病毒增殖的作用.本试验结果为鸭Viperin蛋白抗病毒生物学功能的进一步研究奠定了良好基础.