稳定表达山羊SLAM受体的Vero细胞系的建立  

Establishment of Vero Cell Line Expressing Goat SLAM Receptor Stably

作　　者：高华峰[1] 赵文华[1] 严欢[2] 杨仕标[1] 

机构地区：[1]云南省热带亚热带动物病毒病重点实验室,昆明650224 [2]云南农业大学,昆明650201

出　　处：《中国畜牧兽医》2016年第8期2047-2052,共6页China Animal Husbandry & Veterinary Medicine

基　　金：国家自然科学基金(31160499)

摘　　要：信号淋巴激活分子(signalling lymphocyte activation molecule,SLAM)又称CD150,是小反刍兽疫病毒(peste des petits ruminants virus,PPRV)和犬瘟热病毒(canine distemper virus,CDV)等麻疹病毒属病毒感染淋巴细胞的主要受体,在病毒侵入细胞中发挥着重要作用。为了建立稳定表达山羊SLAM真核细胞系,本研究将全基因合成的gSLAM基因克隆至真核表达质粒pIRES2-GFP中,构建了重组质粒pIRES2-gSLAM。将该重组质粒转染非洲绿猴肾细胞(Vero),经G418筛选后,筛选到稳定表达gSLAM基因的细胞系Vero-gSLAM,该细胞系在传代至第10代,仍能稳定表达gSLAM基因,PPRV N75/1病毒株可以感染且能形成明显的细胞病变(CPE),相比在Vero细胞上10-4.65 TCID50/0.1mL的毒价,在Vero-gSLAM上为10-5.75 TCID50/0.1mL,其毒价有所提高。该细胞系可用于PPRV强毒分离和致弱机制等相关研究。Signalling lymphocyte activation molecule（SLAM,also called CD150）serves as a main cell receptor for PPRV（peste des petits ruminants virus）.This study was aimed to establish a cell line,using Vero cells as the parental cell,to express goat SLAM stably,which could be used to isolate and propagate PPRV.The gene encoding goat SLAMin vitro was synthesized and cloned into eukaryotic expression vector pIRES2-GFP,and the recombinant expression plasmid pIRES2-gSLAM was obtained.The positive stably transfectant Vero-gSLAM cells were screened by G418 and identified by immunofluorescence（IF）and RT-PCR.The result of virus titration by VerogSLAM cell line showed that PPRV strain N75/1had a titre of 10-4.65 TCID50 per 0.1mL in Vero cell at 5day after infection and the titre of PPRV N75/1strain was 10-5.75 TCID50 per 0.1mL in Vero-gSLAM cells.The cell line would play an active role in virus isolation,biological characteristics study and vaccine virus production of PPRV.

关 键 词：小反刍兽疫病毒 受体 信号淋巴细胞激活分子 Vero-gSLAM细胞 

分 类 号：S858.27[农业科学—临床兽医学]