表达猪圆环病毒2型Cap蛋白重组耐热新城疫病毒的构建及免疫原性  

Generation and evaluation of recombinant thermostable Newcastle disease virus expressing Cap protein of porcine circovirus type 2(PCV2)

作　　者：李丽 赵康[1,3] 张媛 王红琳[1] 罗青平[1] 邵华斌[1,2] 潘兹书[3] 温国元 LI Li;ZHAO Kang;ZHANG Yuan;WANG Honglin;LUO Qingping;SHAO Huabin;PAN Zishu;WEN Guoyuan(Institute of Animal Husbandry and Veterinary Sciences,Hubei Academy of Agricultural Sciences,Wuhan 430064,China;Key Laboratory of Prevention and Control Agents for Animal Bacteriosis,Ministry of Agriculture,Wuhan 430064,China;College of Life Science,Wuhan University,Wuhan 430072,China)

机构地区：[1]湖北省农业科学院畜牧兽医研究所,湖北武汉430064 [2]农业部畜禽细菌病防治制剂创制重点实验室,湖北武汉430064 [3]武汉大学生命科学学院,湖北武汉430074

出　　处：《中国兽医学报》2021年第5期865-871,共7页Chinese Journal of Veterinary Science

基　　金：国家重点研发计划资助项目(2017YFD0501104);国家现代农业产业技术体系建设专项资金资助项目(CARS-43-G13);国家自然科学基金资助项目(31873018,31670157);湖北省自然科学基金杰出青年人才资助项目(2019CFA071);湖北省对外科技合作资助项目(2019AHB066)。

摘　　要：猪圆环病毒2型(PCV2)是引起断奶仔猪多系统衰竭综合征(PMWS)的主要病原,给全球养猪业造成了巨大的经济损失。为研发新型PCV2疫苗,采用反向遗传操作技术,以耐热低毒的新城疫病毒TS09-C株为载体,将PCV2主要免疫原蛋白Cap以单独编码框的方式插入至TS09-C株的基因P和M之间,为增强Cap蛋白在载体内的高效表达及细胞外分泌,将Cap基因的核定位信号序列替换为组织纤溶酶原激活物信号肽序列(tPAs)。通过病毒拯救获得了重组新城疫病毒rTS-tPAs-Cap/N。Western blot和间接免疫荧光试验证实,重组病毒可在BHK-21细胞中高效表达Cap蛋白。生物学特性试验表明重组病毒保持了亲本TS09-C株的耐热、低毒和高细胞增殖特性,重组病毒免疫小鼠后可诱导机体产生较高的PCV2抗体水平。本研究为研发耐热、安全、免疫性好的PCV2疫苗奠定了良好的工作基础。Postweaning multisystemic wasting syndrome(PMWS) is one of the most important infectious diseases of swine industry due to the potential for devastating economical losses.Porcine circovirus type 2(PCV2) is the main causative agent of the disease.To develop novel PCV2 vaccine, we constructed recombinant NDV strain rTS-tPAs-Cap/N,based on the thermostable avirulent NDV TS09-C strain to express Cap protein of PCV2.To enhance the expression and extracellular secretion of CAP protein, the nuclear localization signal sequence of CAP gene was replaced by the gene of the tissue plasminogen activator signal peptide(tPAs).The expression of Cap protein could be detected in BHK-21 cells infected with rTS-tPAs-Cap/N virus by Western blot and IFA assays.The recombinant virus retained the thermostability, avirulence and high growth titer of the parental TS09-C virus.Vaccination of mice with the rTS-tPAs-Cap/N virus induced high antibody response to PCV2.Our research provided a basis for the development of thermostable, safe and effective vaccine against PCV2.

关 键 词：猪圆环病毒2型 CAP蛋白 重组新城疫病毒 耐热弱毒株 

分 类 号：S852.65[农业科学—基础兽医学]