[关键词]
[摘要]
为了提高干扰素抗病毒活性测定的生物安全性,本研究使用含有GFP的复制缺陷型水疱性口炎病毒 (VSV△G*G) 为指示病毒,分别对经原核表达系统和杆状病毒表达系统表达的重组猪γ干扰素 (PoIFN-γ) 在MDBK细胞上进行抗病毒活性测定。结果显示:由杆状病毒表达的重组PoIFN-γ具有高度抗病毒活性,其抗病毒活性为105 IU/mL,原核表达的重组PoIFN-γ纯化后经缓慢复性也会产生一定的抗病毒活性,其抗病毒活性为32 IU/mL。该方法与利用表达GFP的重组水疱性口炎病毒 (VSV*GFP) 所检测的干扰素抗病毒活性结果完全一致,表明复制缺陷型病毒VSV△G*G可作为复制型重组病毒的替代品,使得干扰素抗病毒活性检测更加安全、准确。
[Key word]
[Abstract]
In order to ensure the biosafty of the IFN-γ antiviral activity assay, we used a replication-deficient VSV carrying GFP as an interferon sensitive indicator virus (VSV△G*G). The antiviral activities of porcine IFN-γ expressed in Escherichia coli and in baculovirus on MDBK cells were assessed. The results showed that the antiviral activity of porcine IFN-γ expressed in baculovirus could reach 105 IU/mL, while the porcine IFN-γ expressed in E. coli showed some antiviral activity (32 IU/mL) after refolding. The results of the VSV△G*G-based antiviral assay were almost identical to that of the VSV*GFP-based assay, suggesting it is highly feasible to use VSV△G*G as a substitute for VSV*GFP, making assays for IFN-γ antiviral activity safer and more accurate.
[中图分类号]
[基金项目]
国家重点基础研究发展计划 (973计划) (No. 2005CB523202) 资助。