猪圆环病毒1型感染性克隆的构建  

Construction of an infectious clone of porcine circovirus type 1

作　　者：邵小雪[1,2] 周双海[1] 谢俊岭[1] 冉多良[2] 

机构地区：[1]北京农学院动物科学技术学院,北京102206 [2]新疆农业大学动物医学院,新疆乌鲁木齐830000

出　　处：《中国兽医科学》2011年第2期148-151,共4页Chinese Veterinary Science

基　　金：北京市教委科技计划项目(KM200810020013);北京市自然科学基金项目(6082008)

摘　　要：为了构建猪圆环病毒1型(PCV1)感染性DNA克隆,并为PCV1基因突变体的构建及PCV2的相关研究奠定基础,采用PCR方法从PCV1质粒中扩增出了PCV1全基因组片段,将其插入到pBluescriptⅡSK(+)载体中,构建了单拷贝PCV1DNA克隆,并进一步构建了双拷贝PCV1DNA克隆;将双拷贝PCV1DNA克隆转染入PK-15细胞以获得拯救病毒,并初步测定了拯救病毒在体外细胞培养的增殖能力和遗传稳定性。结果表明,成功获得了PCV1拯救病毒,并能够在PK-15细胞中进行稳定传代,其TCID50在连传5代后达到106.55/mL,与其亲本病毒相近,显示出较高的增殖能力。The objective of the present study was to construct the infectious DNA clone of porcine circovirus type 1（PCV1） and lay the foundation for studies of the PCV1 and PCV2 mutant construction.The complete genome of PCV1 was amplified from the PCV1 plasmid by PCR,and was inserted into the pBluescriptⅡSK（＋） vector.After the single copy of PCV1 DNA clone was constructed,then the double copy of PCV1 DNA clone was developed.To obtain the rescue viruses of PCV1,the double PCV1 DNA clone was transfected into porcine kidney（PK）-15 cells.The reproducibility and genetic stability of the viruses were detected by cell culture in vitro.The results indicated that the rescue viruses of PCV1 were obtained successfully and could propagate stably in PK-15 cells,its 50% tissue culture infectious doses was 106.55/mL after 5 serial passages,which was close to its parent and revealed its relatively high infectious titer.

关 键 词：猪圆环病毒1型 感染性DNA克隆 拯救病毒 

分 类 号：S852.659.2[农业科学—基础兽医学]