非结构蛋白2高变区在PRRSV复制中的作用  

Role of PRRSV nsp2 middle hyper variable region playing in the virus life circle

作　　者：吕健[1,2] 韦祖樟[1] 高飞[1] 郑海红[1] 童光志[1] 袁世山[1] 

机构地区：[1]中国农业科学院上海兽医研究所,上海200241 [2]国家知识产权局专利局专利审查协作北京中心,北京100193

出　　处：《中国兽医科学》2012年第10期991-997,共7页Chinese Veterinary Science

基　　金：国家自然科学基金项目(30972204);青年科学基金项目(30901078)

摘　　要：为了研究nsp2的非必需区在动脉炎病毒复制中的作用,采用定点突变PCR(site direct PCR)法对PRRSV全长感染性克隆pAPRRS的nsp2进行了系列缺失。结果显示,通过病毒拯救发现,nsp2部分区段可以忍受一定程度的缺失,但大幅度的缺失影响了病毒的复制、RNA合成翻译甚至病毒颗粒的成熟过程。结果表明,nsp2中多个氨基酸的变化可能影响PRRSV中多聚蛋白的水解、加工、成熟或RTC的组装。In order to research the importance of different regions of nsp2 in the life cycle of porcine reproductive and respiratory syndrome virus（PRRSV） in vitro,a series of mutants containing in-frame deletion in the nsp2-coding region based on the full length infectious clone of pAPRRS were engineered.In result,some infectious viruses were rescued from transfected cells and further identified.The total PL2（47-147 aa） and the putative transmembrane domain were essential for replication.On the contrast,the nsp2 middle hyper variable region（148-911 aa） can tolerate 100aa or more amino acids deletions but could not be removed completely;the largest deletion was about 199 aa（nsp331-529）.The rescued virus exhibited the similar CPE,growth kinetics and mRNA transcription in the transfected MA-104 cells.In conclusion,some part of the nsp2-coding region was essential for productive virus infection,and partial deletion can be tolerated in the life cycle of PRRSV in cell culture.

关 键 词：猪繁殖与呼吸障碍综合征病毒 亲水区 跨膜区 病毒复制 

分 类 号：S852.659.6[农业科学—基础兽医学]