猪圆环病毒2型ORF2和猪繁殖与呼吸综合征病毒ORF5重组杆状病毒疫苗的研究  

The Study on Recombinant Baculovirus Vaccine of PCV2 ORF2 and PRRSV ORF5

作　　者：张翔峰[1] 何逸民[1] 曹宗喜[1] 孔留五[1] 张得玉[1] 秦宏阳[1] 张桂红[1] 

机构地区：[1]华南农业大学兽医学院,广州510642

出　　处：《中国畜牧兽医》2010年第9期62-67,共6页China Animal Husbandry & Veterinary Medicine

基　　金：国家现代生猪现代农业产业技术体系项目(nycytx009);家畜重要病毒基因工程疫苗研究和创制(2006AA10A204);国家高技术研究发展计划(863计划)项目

摘　　要：参照GenBank上收录的基因组序列自行设计引物,分别扩增猪圆环病毒2型ORF2和猪繁殖与呼吸综合征病毒ORF5。将外源基因分别插入经改造的杆状病毒穿梭载体pFVC,获得3种重组质粒pFVC-ORF2、pFVC-ORF5、pFVC-ORF5-ORF2。将重组质粒pFVC-ORF2、pFVC-ORF5、pFVC-ORF5-ORF2分别转入带有杆状病毒骨架的DH10BacTME.coli感受态细胞中。提取高质量的重组质粒rBacmid-ORF2、rBacmid-ORF5、rBacmid-ORF5-ORF2,将重组质粒转染Sf9细胞,获取重组病毒rAc-ORF2、rAc-ORF5、rAc-ORF5-ORF2。通过间接免疫荧光检测证实了外源基因正确表达。重组杆状病毒rAc-ORF2、rAc-ORF5、rAc-ORF5-ORF2对BALB/c小鼠的免疫试验结果显示,rAc-ORF2、rAc-ORF5、rAc-ORF5-ORF2均能够刺激小鼠机体产生免疫应答,产生抗体,并能持续较长时间。Several pairs of primers were designed according to the genome sequences,primers were designed to amplify PCV2 ORF2 and PRRSV ORF5.The foreign gene were inserted into the modified baculovirus shuttle vector pFVC,three kinds of recombinant plasmid pFVC-ORF2,pFVC-ORF5,pFVC-ORF5-ORF2 were successfully obtained,and the recombinant plasmid pFVC-ORF2,pFVC-ORF5,pFVC-ORF5-ORF2 were respectively transformed into DH10BacTM E.coli which contain baculovirus skeleton.Then high-quality recombinant rBacmid-ORF2,rBacmid-ORF5,rBacmid-ORF5-ORF2 were extracted,and transfect to Sf9 cell.Three kinds of recombinant baculovirus rAc-ORF2,rAc-ORF5,rAc-ORF5-ORF2 were successfully rescued.The foreign protein were detected by IFA.Recombinant baculovirus rAc-ORF2,rAc-ORF5,rAc-ORF5-ORF2 were injected into BALB/c mice for immunization experiments,results showed that rAc-ORF2,rAc-ORF5,rAc-ORF5-ORF2 were able to stimulate the body immune response,produce antibodies,and sustain for a long time.

关 键 词：猪圆环病毒2型 猪繁殖与呼吸综合征病毒 重组杆状病毒 BALB/c小鼠免疫试验 

分 类 号：S852.65[农业科学—基础兽医学]