鸭坦布苏病毒E基因DNA疫苗构建及免疫原性的初步研究  

Construction and immunogenicity of DNA vaccine encoding E gene of duck Tembusu virus

作　　者：徐大伟[1,2] 李国新[2] 李雪松[2] 姬希文[1,2] 李云章[1] 李泽君[2,3] 

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]中国农业科学院上海兽医研究所农业部寄生虫学重点开放实验室,上海200241 [3]国家家禽工程技术研究中心,上海200241

出　　处：《中国预防兽医学报》2012年第4期305-308,共4页Chinese Journal of Preventive Veterinary Medicine

基　　金：公益性行业(农业)科研专项经费项目(201003012);自然科学基金面上项目(31172332)

摘　　要：鸭坦布苏病毒(DTMUV)感染引起产蛋鸭产蛋急剧下降。目前,没有预防该病的疫苗。为研究DTMUV DNA疫苗的免疫效果,本研究将DTMUV的E基因插入pCAGGS载体,构建了重组质粒pCAGGS-E。将pCAGGS-E转染293T细胞后,采用间接免疫荧光(IFA)和western blot检测E蛋白的表达情况。结果显示,两种方法均检测到了E蛋白的特异性表达。将pCAGGS-E用脂质体包裹后通过尾静脉注射免疫BALB/c小鼠,采用间接ELISA检测抗体的产生情况。结果表明,随着免疫次数的增加及免疫时间的延长,免疫小鼠的抗体滴度逐渐升高。本研究证明,编码E基因的重组质粒DNA免疫小鼠后能够诱导有效的免疫应答,为DTMUV DNA疫苗的研究奠定了基础。Duck Tembusu virus（DTMUV） infection leads to decrease dramatically in egg production of laying duck.However,no vaccine against DTMUV is available at present.To investigate the immunogenicity of DNA vaccine against DTMUV,the E gene of DTMUV was cloned into eukaryotic expression vector pCAGGS to construct pCAGGS-E and the expression of E protein was detected in pCAGGS-E transfected 293T cells by indirect immunofluorescence assay and western blot.Furthermore,the immunogenicity was evaluated in BALB/c mice with the liposome capsuled pCAGGS-E via intravenous injection.The specific antibodies were detected by indirect ELISA,which showed the antibody titers increased gradually after pCAGGS-E multiple inoculations.In conclusion,the pCAGGS-E was able to induced specific immune responses in mice.

关 键 词：鸭坦布苏病毒 DNA疫苗 E基因 

分 类 号：S852.65[农业科学—基础兽医学]