猪细小病毒结构蛋白VP_1和VP_2的基因免疫研究  

Study on Gene Immunity of Porcine Parvovirus Structural Protein Genes VP_1 and VP_2

作　　者：赵俊龙[1] 陈焕春[1] 吕建强[1] 王祥[1] 肖少波[1] 周复春[1] 

机构地区：[1]华中农业大学畜牧兽医学院,湖北武汉430070

出　　处：《病毒学报》2003年第1期47-51,共5页Chinese Journal of Virology

基　　金：国家九五重中之重课题 ( 96 -0 0 3 -0 1-0 41);湖北省自然科学基金 ( 99J10 0 )资助。

摘　　要：利用真核表达载体pCIneo和pcDNA3 1( +)分别构建了含有猪细小病毒VP1基因的 pCIneoVP1和含有VP2基因的 pCIneoVP2 与 pcDNAVP2 三种真核表达质粒。将上述三种真核表达质粒分别转染IBRS 2细胞 ,利用间接ELISA检测表达情况 ,结果表明上述三种质粒均能在IBRS 2细胞表达 ,表达产物位于细胞中。在此基础上 ,利用这三种质粒分别以肌内注射的方式 ,间隔 2周 2次免疫小鼠 ,结果发现所有表达质粒均能诱导产生明显的细胞免疫和体液免疫 ,其中pCIneoVP1质粒诱导的体液免疫最强 ,与猪细小病毒灭活疫苗免疫组相当 ,pCIneoVP2 诱导的细胞免疫应答强于PPV灭活组 ,pCIneoVP1和 pCIneoVP2 联合免疫并没有加强作用。The complete VP 1 and VP 2 genes were cloned into eukaryotic expression vectors of pcDNA 3.1(+) and pCIneo,resulting the recombinant expression plasmids pcDNAVP 2,pCIneoVP 2 and pCIneoVP 1.Then these plasmids were separately transfected into IBRS-2 cell and several clone cells were obtained under the selection of G418.The expressed protein was detected by ELISA.On the basis of above works,these plasmids were used as DNA vaccine to immunize the mice.The results showed that all these plasmids could induce specific cell-mediated and humoral immunity.The specific cell-mediated immunity induced by pCIneo VP 2 was stronger than that induced by the inactivated PPV vaccine,the humoral immunity induced by plasmid pCIneo VP 1 was similar with that induced by inactived PPV vaccine.The immune responses induced by co-immunization with pCIneo VP 1 and pCIneo VP 2 were not stronger than that induced by pCIneo VP 2 or pCIneo VP 1 alone.These preliminary results show the hope of developing PPV DNA vaccine and the study is worthy to continue.

关 键 词：猪细小病毒 结构蛋白 真核表达 基因免疫 

分 类 号：S852.659.2[农业科学—基础兽医学]