尼帕病毒F糖蛋白在重组牛痘病毒中的表达及鉴定

Expression of Nipah Virus Fusion Protein in Recombinant Vaccinia Virus and Assays of Its Biological Function

尼帕病毒(NiV)F蛋白在病毒侵入细胞和诱导中和抗体等方面具有重要作用。通过over-lapping PCR合成密码子优化的F蛋白基因构建了表达NiV F蛋白的重组牛痘病毒(WR株)rWR-NiV-F。利用兔抗NiV血清为检测抗体,通过间接免疫荧光(IFA)检测到了F蛋白在重组病毒感染细胞中的表达。SDS-PAGE和Western blot检测证明重组蛋白F0被裂解为F1和F2。以rWR-NiV-F感染瞬时转染共表达NiV受体结合囊膜糖蛋白G的BHK细胞,可诱导细胞膜融合及合包体形成,证明该重组病毒表达F蛋白保持良好的抗原性及生物学活性,为NiV诊断及重组活载体疫苗研究奠定了重要基础。

The envelope fusion glycoprotein （F） of Nipah virus （NiV） plays key role in viral entry and eliciting neutralization antibody, In this study, the mammalian condon optimized F gene was synthesized by over-lapping PCR and used to generate recombinant vaccinia virus, rWR-NiV-F. The expression of Nipah virus F protein in rWR NiV F infected cells was detected by indirect immunoflorescence assay with rabbit serum anti-Nipah virus. The efficient cleavage of F protein expressed by rWR-NiV-F was also confirmed by SDS-PAGE and Western blot, Furthermore, syncytium formation could be induced in BHK cells by rWR-NiV-F infection following transfection of NiV G protein expression plasmid. The results demonstrated the fusion protein expressed by rWR-NiV-F keeps native antigenicity and biological fusion activity.