以rLaSota为骨架构建表达强毒株HN蛋白的嵌合体新城疫病毒  

Construction of Newcastle disease virus chimeras expressing the hemagglutinin-neuraminidase protein of velogenic strain F48e9 using rLaSota as backbone

作　　者：丁玉林[1,2] 王永[1] 解希帝[1] 葛金英[2] 步志高[2] 王凤龙[1] 

机构地区：[1]内蒙古农业大学兽医学院,内蒙古呼和浩特010018 [2]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,黑龙江哈尔滨150001

出　　处：《畜牧与兽医》2010年第11期1-6,共6页Animal Husbandry & Veterinary Medicine

基　　金：国家科技攻关项目(2004BA519A19;2005BA711A10);"973项目"(2005CB523200)

摘　　要：新城疫病毒(NDV)的HN蛋白是一个多功能蛋白,具有血凝素(HA)和神经氨酸酶(NA)两种活性,在病毒感染过程中扮演重要角色。本研究利用反向遗传操作技术,将NDV强毒株F48E9的HN基因替换弱毒株rLaSota的HN基因,获得嵌合病毒rL-F48E9HN。收获病毒尿囊液,提取基因组RNA,进行序列分析,结果显示嵌合病毒基因组的HN基因获得了正确替换。嵌合病毒在鸡胚内的生长特性与亲本株LaSota一致,与F48E9的生长特性相差甚远。嵌合病毒红细胞吸附性明显增强,较rLaSota株增加51%,而细胞融合作用无明显变化。rL-F48E9HN的鸡胚平均致死时间(MDT)为148 h,脑内致病指数(ICPI)为0.43,静脉接种致病指数(IVPI)为0,说明rL-F48E9HN的毒力仍然属于弱毒力范围,而没有达到中等毒力或者强毒力。The hemagglutinin-neuraminidase （HN） protein of Newcastle disease virus （NDV） plays a crucial role in the process of infec- tion. In this study, we exchanged the HN gene of the velogenic strain by means of reverse genetics procedures. The HN gene of the attenua- ted recombinant NDV strain rLaSota was replaced by the HN gene of the virulent strain F48E9 to obtain the chimeric virus rL-F48E9HN. The aUantoic fluid from the embryonated chicken eggs infected with the rescued virus rL-F48E9HN was harvested. Sequence analysis of viral ge- nomic RNA showed that the HN gene in the genome of chimeric virus was correctly replaced. Growth characteristics of the chimeric virus in embryonated specific-pathogen-free （SPF） chicken eggs were consistent with the parent strain rLaSota, but far from F48E9. Chimeric virus increased by 51% in hemadsorption activity, compared with rLaSota strains, while no significant change occurred in cell fusion. The embryo mean death time （MDT） of rL-F48E9HN is 148 h, the intracerebral pathogenicity index （ICPI） is 0.43, and the intravenous pathogenicity index （IVPI） is 0. It suggested the chimeric virus rL-F48E9HN still belonged to the attenuated strain, not moderately or highly virulent strain.

关 键 词：新城疫病毒 HN蛋白 弱毒株rLaSota 强毒株F48E9 反向遗传操作 

分 类 号：R392[医药卫生—免疫学]