狂犬病病毒糖蛋白跨膜区和膜内区的替换对其生物学特性的影响  

The impact of the replacement of transmembrane domain and cytoplasmic tail of rabies virus glycoprotein on its biological characteristics

作　　者：于桂梅[1] 周微微[1] 祖述龙 王子龙[1,2] 陈曦[1] 葛金英[1] 步志高[1] 

机构地区：[1]中国农业科学院哈尔滨兽医研究所兽医生物技术国家重点实验室/农业部兽医公共卫生重点实验室,黑龙江哈尔滨150001 [2]河北农业大学动物医学学院,河北保定071000

出　　处：《中国预防兽医学报》2015年第8期571-575,共5页Chinese Journal of Preventive Veterinary Medicine

基　　金：中国国家科技计划(2012ZX10004214)

摘　　要：为研究表达狂犬病病毒(RV)糖蛋白(G)重组新城疫病毒(r L-RVG)中RV G跨膜区(TM)及膜内区(CT)对RV G蛋白表达、病毒蚀斑的形成以及免疫原性的影响,本研究采用新城疫病毒(NDV)融合蛋白(F)的TM和CT替换G蛋白的相应部分,构建出表达RV嵌合G蛋白(RVGTC)的重组NDV(r L-RVGTC)。激光共聚焦和western blot检测结果显示,r L-RVG和r L-RVGTC在BHK-21细胞中RVG表达量无显著差异;而且r L-RVG与r L-RVGTC在鸡胚中的病毒增殖滴度也无显著差异。但间接免疫荧光结果表明,r L-RVGTC在BHK-21细胞中丧失RV或r L-RVG形成病毒蚀斑的能力。并且以5×107半数鸡胚感染量(EID50)的r L-RVG和r L-RVGTC分别免疫小鼠后,r L-RVGTC诱导产生的RV中和抗体的滴度显著低于r L-RVG诱导的滴度。本实验结果显示TM和CT对RV G的病毒蚀斑形成及其免疫原性是必不可少的。In order to tudy the impact of replacement of the transmembrane domain （TM） and cytoplasmic tail （CT） on the rabies virus （RV） glycoprotein protein （G） expression, the formation of virus plaque and immunogenicity of recombinant NDV LaSota strain （rL-RVG）, the recombinant virus of rL-RVGTC was constructed, expressing a chimeric RV G protein based on rL-RVG, in which the TM and CT of RVG were replaced with that of the NDV fusion （F） glycoprotein. The chimeric RV G protein of RVGTC was abundantly expressed at the surface of infected BHK-21 cells in a similar level with rL-RVG, but failed to produce the plaques in the cell culture. In addition, rL-RVG and rL-RVGTC virus grew to similar titers in chicken embryo. However, in mice, the RV neutralizing antibody inducted by rL-RVGTC in mice was significantly lower than that by rL-RVG. These results indicated that the TM and CT of RVG were essential for the formation of virus plaque and its immunogenicity.

关 键 词：狂犬病病毒 G蛋白 新城疫病毒 F蛋白 跨膜区 膜内区 

分 类 号：S852.65[农业科学—基础兽医学]