中国兽医科学2017，Vol.38Issue(8)：1022-1026,5.DOI:10.16656/j.issn.1673-4696.2017.08.015
小反刍兽疫病毒受体细胞系的构建及应用
Construction and application of receptor cell line of peste des petits ruminants virus
摘要
Abstract
To establish cell lines with a stable expression of signaling lymphocyte activation molecule of goat,and to research the PPRV infection mechanisms,separation of virus and vaccine production,the extracellular domain of SLAM gene was amplified by PCR,then inserted into the pDisplay vector and named pDisplay-gSLAM.G418 and anti-SLAM monoclonal antibody were used to screen theVero-E6 cell lines that could stably express gSLAM.The cell lines were identified by PCR,indirect immunofluorescence test and Western-blot.At the same time,PPRV virus was isolated by this cell line,and this cell line was absorbed by the PPRV isolate and vaccine strains together with Vero-E6 cells.In result,gSLAM/Vero-E6 was successfully screened,and the gSLAMgene was successfully amplified by PCR method.The gSLAM gene was successfully expressed on the cell membrane by indirect immunofluorescence and Western-blot,and the PPRV was isolated.At the same time,the adsorption of PPRV isolates and vaccine strains,compared with Vero-E6 cells,gSLAM/Vero-E6 cell lines had more obvious pathological changes.In conclusion,gSLAM/Vero-E6 cells are highly sensitive to PPRV in clinical specimens and provides experimental materials for follow-up studies of PPRV.关键词
小反刍兽疫病毒/pDisplay载体/细胞系Key words
peste des petits ruminants virus/pDisplay vector/cell line分类
农业科技引用本文复制引用
金红岩,封家旺,梁琳,赵玲娜,隋修锟,史利军,侯绍华,李刚..小反刍兽疫病毒受体细胞系的构建及应用[J].中国兽医科学,2017,38(8):1022-1026,5.基金项目
国家自然科学基金资助项目(31172342) （31172342）
国家科技支撑计划项目(2013BAD12B05) （2013BAD12B05）
国际合作项目(D32025/17453) （D32025/17453）
