羊干扰素α和γ融合表达及抗病毒活性分析  

Fusion expression and biological activity of sheep interferonαandγ

作　　者：韩颖[1] 李秀丽[1] 豆子航 郭笑然 雷白时[1] 袁万哲[1,2] 赵款 HAN Ying;LI Xiuli;DOU Zihang;GUO Xiaoran;LEI Baishi;YUAN Wanzhe;ZHAO Kuan(College of Veterinary Medicine,Hebei Agricultural University,Baoding,Hebei 071001,China;Hebei Veterinary Biotechnology Engineering Technology Innovation Center,Baoding,Hebei 071001,China)

机构地区：[1]河北农业大学动物医学院,河北保定071001 [2]河北省兽医生物技术工程技术创新中心,河北保定071001

出　　处：《中国兽医学报》2023年第4期660-667,共8页Chinese Journal of Veterinary Science

基　　金：河北省现代农业产业技术体系羊产业创新团队专项资助项目(HBCT2018140204);河北农业大学精准畜牧学科群建设资助项目(1090064)。

摘　　要：为进一步探究羊干扰素α和γ的抗病毒活性效果,根据GenBank羊IFN-α和IFN-γ基因序列,使用Linker连接合成目的序列,成功构建了3种重组表达质粒pET28a-OviIFN-γ、pET28a-OviIFN-α、pET28a-OviIFN-γ/α并进行原核表达。用细胞病变抑制法和RT-qPCR测定重组蛋白rOviIFN-α、rOviIFN-γ和rOviIFN-α/γ的抗病毒活性。结果表明,rOviIFN-α、rOviIFN-γ和rOviIFN-α/γ在牛肾细胞(MDBK)以及非洲绿猴肾细胞(Vero)细胞中有抗病毒活性,且rOviIFN-α/γ的抗病毒效果最优;rOviIFN-α、rOviIFN-γ和rOviIFN-α/γ能够显著上调MDBK以及Vero细胞中4种干扰素刺激基因(IFN stimulated genes,ISGs)的mRNA转录水平,且rOviIFN-α/γ对干扰素刺激基因的上调水平最显著。综上所述,本试验表达的重组蛋白都具有抗病毒作用,其中串联表达的重组蛋白抗病毒活性以及干扰素刺激基因转录水平较高。In order to explore the antiviral activity of recombinant sheep interferon,the gene sequences of IFN-αand IFN-γrecorded in GenBank were synthesized by linker ligation.Three recombinant expression plasmids,pET28a-OviIFN-γ、pET28a-OviIFN-α、pET28a-OviIFN-OviIFN-α/γwere successfully constructed and transformed into Escherichia coli BL21(DE3)for prokaryotic expression.The antiviral activities of rOviIFN-α,rOviIFN-γand rOviIFN-α/γwere determined by cytopathic inhibition and RT-qPCR.The recombinant fusion protein has antiviral activity in MDBK and Vero cells.The recombinant fusion protein could significantly upregulate the mRNA transcription levels of four interferon stimulating genes(ISGs)in MDBK and Vero cells.In conclusion,the recombinant proteins expressed in this experiment have antiviral effect,with the recombinant proteins expressed in tandem having higher antiviral activity as well as interferon-stimulated gene transcription levels.

关 键 词：羊干扰素α和γ 细胞病变抑制 融合表达 抗病毒活性 干扰素刺激基因 

分 类 号：S825.65[农业科学—畜牧学]