嵌合型PCV1-3感染性克隆的构建及病毒拯救  

Construction and Virus Rescue of Chimeric PCV1-3 Infectious Clone

作　　者：李佳昕 田瑶 韩先杰[1] 杨莹 王硕 时建立[2] 李琛 刘畅[2] 彭喆[2] 吴晓燕[2,3] 韩红 李俊[1,2,3] LI Jiaxin;TIAN Yao;HAN Xianjie;YANG Ying;WANG Shuo;SHI Jianli;LI Chen;LIU Chang;PENG Zhe;WU Xiaoyan;HAN Hong;LI Jun(College of Veterinary Medicine,Qingdao Agricultural University,Qingdao,Shandong 266109,China;Institute of Animal Science and Veterinary Medicine/Shandong Academy of Agricultural Sciences,Ji’nan,Shandong 250100,China;College of Life Science,Shandong Normal University,Ji’nan,Shandong 250014,China)

机构地区：[1]青岛农业大学动物医学院,山东青岛266109 [2]山东省农业科学院畜牧兽医研究所,山东济南250100 [3]山东师范大学生命科学院,山东济南250014

出　　处：《家畜生态学报》2023年第10期77-81,96,共6页Journal of Domestic Animal Ecology

基　　金：山东省重大科技创新工程(2019JZZY010720);山东省现代农业产业技术体系疫病控制岗位(SDAIT-08-07);泰山学者项目。

摘　　要：为了构建猪圆环病毒1型(porcine circovirus type 1,PCV1)和猪圆环病毒3型(porcine circovirus type 3,PCV3)的嵌合病毒PCV1-3,并为PCV3疫苗研制奠定基础,该研究以PCV1基因组为骨架,将PCV1的ORF2基因替换为PCV3的ORF2基因,构建嵌合型猪圆环病毒(PCV1-3)DNA克隆。PCR和序列测序结果显示,成功构建出了PCV1-3感染性克隆;将感染性克隆转染PK-15细胞,并将第4代病毒进行免疫过氧化物酶单层试验(IPMA)和蛋白免疫印迹(Western Blot)检测,结果显示成功拯救了重组病毒PCV1-3。对第4代病毒进行滴度测定,效价为105.13 TCID 50/mL。因此,该试验成功拯救出一种新型嵌合型病毒PCV1-3,并为进一步研究PCV3疫苗奠定了基础。In order to construct porcine circovirus type 1(PCV1)and porcine circovirus type 3(PCV3)chimeric viruses PCV1-3,and to lay the foundation for the development of PCV3 vaccine,this study constructed a chimeric porcine circovirus(PCV1-3)DNA clone by replacing the ORF2 gene of PCV1 with the ORF2 gene of PCV3 using the PCV1 genome as the skeleton.PCR and sequencing results showed that the infected clone of pCv1-3 was successfully constructed.The infected clone was transfected into PK-15 cells,and the 4th generation virus was detected by immunoperoxidase monolay assay(IPMA)and Western Blot,and the recombinant virus PCV1-3 was successfully rescued.The titer of the fourth-generation virus was measured and the titer was 105.13 TCID 50/mL.A new chimeric virus PCV1-3 was successfully rescued and it laid a foundation for further research on PCV3 vaccine.

关 键 词：嵌合型病毒 感染性克隆 PCV1 PCV3 

分 类 号：S852.65[农业科学—基础兽医学]