鸡源ADAR2基因克隆及其抗新城疫病毒作用研究

摘要

目前对家禽中双链RNA特异性腺苷脱氨酶(adenosine deaminase acting on double-stranded RNA,ADAR)的种类、组织分布及其功能尚不明了.本研究中以刀豆素、poly I:C以及新城疫病毒(Newcastle disease virus,NDV)作为刺激物诱导鸡胚成纤维细胞(chicken embryo fibroblast,CEF)中ADARs基因上调,扩增获得鸡ADAR2基因的mRNA.为了鉴定ADAR2的抗病毒作用,本研究构建了能够偶联有绿色荧光蛋白(green fluorescent protein,GFP)的ADAR2重组真核表达质粒,转染DF1细胞,结果显示在过量表达ADAR2的细胞中,NDV的增殖受到了显著抑制,表明ADAR2在NDV感染过程中发挥重要的抗病毒作用,这为进一步研究鸡RNA编辑酶对NDV RNA编辑奠定了基础.%The type, tissue distribution and function of avian adenosine deaminase acting on double-stranded RNA (ADAR) have not been well characterized although recently it has been extensively studied for its roles in antiviral effect and virus evolution. In this study, ADAR2 mRNA was amplified from primary CEF cells previously stimulated with ConA, poly I:C and Newcastle disease virus (NDV). The eukaryotic plasmid expressing green fluorescent protein (GFP)-labeled ADAR2 was constructed to determine the antiviral effect of ADAR2. In the transfected DF1 cells, the GFP-labeled ADAR2 was visualized in cellular nuclei and growth of NDV was obviously inhibited, suggesting chicken ADAR2 might play a role in the anti-NDV activity. However, the detailed mechanism of ADAR2 mediated anti-NDV effect need to be carried out in the future.