摘要
为了解猪源IFN-λ3的体外抗病毒活性,本研究利用RT-PCR技术从猪肺巨噬细胞(CRL2845)中扩增获得猪源IFN-λ3基因(pIFN-λ3),并将pIFN-λ3基因克隆至真核表达载体pcDNA3.1,构建重组质粒pCDNA3.1-pIFN-λ3,利用脂质体转染293细胞。48 h后通过Western blot分析pIFN-λ3瞬时表达情况,取转染后上清分别作用于PK15、MDCK细胞进行pIFN-λ3抗病毒活性分析。结果显示,本研究成功扩增获得pIFN-λ3基因,并在293细胞内成功表达;瞬时表达能够抑制表达绿色荧光蛋白的重组水泡性口炎病毒(Vesicular stomatitis virus-green fluorescent protein,VSVG)在PK15、MDCK细胞上的增殖。本研究结果为深入研究猪源IFN-λ3在宿主抗病毒免疫和黏膜免疫中的作用机理及其应用提供参考和依据。
To analyze the i antiviral activity of porcine IFN-λ3 in vitro.The porcine IFN-λ3 gene(pIFN-λ3)was amplified from porcine pulmonary macrophage cells(CRL2845)by RT-PCR.The pIFN-λ3 gene was cloned into eukaryotic expression vector pcDNA3.1 and the recombinant plasmid pCDNA3.1-pIFN-λ3 was constructed,then 293 cells were transfected with liposome.After 48 h,the transient expression of pIFN-λ3 was analyzed by Western blot.The pIFN-λ3 antiviral activity assay was performed on PK15 and MDCK cells.The results show that the pIFN-λ3 gene was successfully amplified and expressed in 293 cells.Transient expression of the gene could inhibit the proliferation of VSVG on PK15 and MDCK cells.The research will lay a foundation for further study on the mechanism and application of porcine IFN-λ3 in host antiviral immunity and mucosal immunity.To analyze the in vitro antiviral activity of porcine IFN-λ3.The porcine IFN-λ3 gene(pIFN-λ3)by porcine pulmonary macrophage(CRL2845)by RT-PCR.pIFN-λ3 gene The recombinant plasmid pCDNA3.1-pIFN-λ3 was cloned into eukaryotic expression vector pcDNA3.1,and 293 cells were transfected with liposome.After 48 h,the transient expression of pIFN-λ3 was analyzed by Western blot.The pIFN-λ3 antiviral activity assay was performed on PK15 and MDCK cells.The pIFN-λ3 gene was successfully amplified and successfully expressed in 293 cells.Transient expression could inhibit the proliferation of VSVG on PK15 and MDCK cells.To lay a foundation for further study on the mechanism and application of porcine IFN-λ3 in host antiviral immunity and mucosal immunity.
作者
陈竞
许汪
宋利娜
郝鹏飞
姜宇航
付婷婷
金鑫
金宁一
李昌
CHEN Jing;XU Wang;SONG Li-na;HAO Peng-fei;JIANG Yu-hang;FU Ting-ting;JIN Xin;JIN Ning-yi;LI Chang(Yanbian University Agricultural College,Yanji 133000,China;Military Institute of Military Veterinary Medicine,Academy of Medical Sciences,Changchun 130122,China)
出处
《中国动物传染病学报》
CAS
北大核心
2019年第5期32-37,共6页
Chinese Journal of Animal Infectious Diseases
基金
国家重点研发计划资助项目(2017YFD0501002)
国家自然科学基金(31702210)
