中国兽医科学2025，Vol.55Issue(4)：456-462,7.DOI:10.16656/j.issn.1673-4696.2025.0072
稳定表达小反刍兽疫病毒N蛋白的THP-1细胞系的建立与应用
Establishment and application of THP-1 cell line with stable expression N protein of peste des petits ruminants virus
摘要
Abstract
The aim of this study was to construct a THP-1 cell line that stably express the N protein of peste des petits ruminants virus(PPRV)by using a lentiviral vector.And the effect of this cell line on the inflammatory response activated by PPRV was identified.Firstly,the N gene sequence was amplified through PCR,and then the lentivirus expression vector pCDH-CMV-MCS-EF1-CopGFP-T2A-Puro-N was constructed following double-enzyme digestion.Subsequently,the vector and lentivirus packaging plasmids,namely pLP1,pLP2 and pLP/VSVG,were co-transfected into HEK293T cells to obtain lentivirus capable of expressing the N protein.Furthermore,the THP-1 cells were infected with this lentivirus,and then screened with puromycin.Then the THP-1 cell line with stable expression of the N protein(THP-1-N)was successfully obtained by limited dilution method.Finally,ELISA and Western-blot were used to detect the effect of this cell line on the inflammatory response activated by PPRV.The results showed that the THP-1-N cell line was successfully constructed,which not only had good viability,but also had the ability of stable passage.Besides,the results demonstrated that THP-1-N cells could markedly promote the expression of IL-1β in-duced by PPRV and trigger an inflammatory response.Taken together,THP-1 cell line with stable expression of PPRV N protein was successfully established,which provided important biological materials for the study of PPRV infection mechanism.关键词
小反刍兽疫病毒/N蛋白/慢病毒/细胞系/炎症小体Key words
peste des petits ruminants virus/N protein/lentivirus/cells line/inflammasome分类
农业科技引用本文复制引用
李玲霞,张旭辉,令小东,韩生义,吴锦艳,尚佑军..稳定表达小反刍兽疫病毒N蛋白的THP-1细胞系的建立与应用[J].中国兽医科学,2025,55(4):456-462,7.基金项目
2024年度宁夏回族自治区重点研发计划项目(2024BBF02021) （2024BBF02021）
省部共建三江源生态与高原农牧业国家重点实验室自主课题(2024-ZZ-05) （2024-ZZ-05）
