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PCV2a和PCV2b感染性克隆的体外拯救
引用本文:柳晓,钟辉,孙玉霞,李谦,邹榕凯,李俊,时建立,丛晓燕,孙文博,杜以军,吴家强,王金宝.PCV2a和PCV2b感染性克隆的体外拯救[J].家畜生态学报,2012,33(6):67-71.
作者姓名:柳晓  钟辉  孙玉霞  李谦  邹榕凯  李俊  时建立  丛晓燕  孙文博  杜以军  吴家强  王金宝
作者单位:青岛农业大学动物科技学院;山东省农业科学院;国家知识产权局专利局专利审查协作北京中心
基金项目:山东省自然科学基金资助项目(2010ZRC02031);国家成果转化项目(2011GB2C600011,2010GB2C600259)
摘    要: 研究旨在获得PCV2a和PCV2b的拯救毒株。对筛选得到的PCV2a和PCV2b感染性克隆分别进行酶切,使酶切得到的全长基因组DNA自身环化并分别转染PK15细胞。经IFA和PCR验证,确认成功拯救出两亚型PCV2病毒。病毒传至第9代时对PCV2a和PCV2b的毒价分别进行测定,结果显示PCV2a的毒价为103.5TCID50/mL, PCV2b的毒价为104.6TCID50/mL。本试验为分型特异性序列与病毒致病力相关性的研究打下了基础.

关 键 词:猪圆环病毒2型  分型特异性序列  自身环化  体外拯救
收稿时间:2012-07-20

Identification of Rescued Viruses from Infectious Clones of PCV2a and PCV2b in Vitro
LIU Xiao,ZHONG Hui,SUN Yu-xia,LI Qian,ZOU Rong-kai,LI Jun,SHI Jian-li,CONG Xiao-yan,SUN Wen-bo,DU Yi-jun,WU Jia-qiang,WANG Jin-bao.Identification of Rescued Viruses from Infectious Clones of PCV2a and PCV2b in Vitro[J].Acta Ecologae Animalis Domastici,2012,33(6):67-71.
Authors:LIU Xiao  ZHONG Hui  SUN Yu-xia  LI Qian  ZOU Rong-kai  LI Jun  SHI Jian-li  CONG Xiao-yan  SUN Wen-bo  DU Yi-jun  WU Jia-qiang  WANG Jin-bao
Institution:1,3(1.College of Animal Science and Technology,Qingdao Agricultural University,Qingdao,Shandong,266109,China; 2.Patent Examination Cooperation Center of The Patent Office,SIPO,Beijing,Beijing,100081,China; 3.Shandong Academy of Agricultural Sciences,Jinan,Shandong,250100,China)
Abstract:The study was designed to get the rescued strains of PCV2a and PCV2b.The screened PCV2a and PCV2b infectious clones were digested, then the genomic DNA was cyclized and transfected into PK15 cells.IFA and PCR indicated that two subtypes of PCV2 were rescued successfully. Virus titers in cell cultures for the 9th passage were determined by a microtitration infectivity assay and recorded as TCID50/mL. PCV2a was 103.5TCID50/mL,and PCV2b was 104.6TCID50/mL. This experiment laid the foundation for the study of the correlation between type specific sequence and virus virulence.
Keywords:porcine circovirus type 2 (PCV2  type specific sequence  the ligation DNA of PCV2  characterizing in vitro
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