猪圆环病毒2型感染性克隆的构建  

Construction of the chimeric infectious DNA clone of type 2 porcine circovirus

作　　者：王明亮[1] 刁有祥[1] 纪巍[1] 孔义波[1] 高继明[1] 

机构地区：[1]山东农业大学动物科技学院,山东泰安271018

出　　处：《中国兽医学报》2011年第3期368-372,共5页Chinese Journal of Veterinary Science

摘　　要：利用PCR方法从临床疑似PMWS症状猪的脏器中扩增出1株猪圆环病毒2型(PCV-2)的全基因组(1 767bp),命名为SDZQ-1,对其基因进行克隆测序并与GenBank中收录的21个PCV-2毒株的全基因组序列进行比较,同源性结果为95.0%～99.7%。将2个SDZQ-1的全基因组序列正向串联连接入PUC19载体质粒中,成功构建了PCV-2双拷贝质粒。体外转染PK-15细胞,盲传5代后,用PCR和间接免疫荧光方法检测,结果构建的PCV-2克隆具有感染性。A full-length genome DNA of PCV-2 was amplified by PCR and sequenced,then named SDZQ-1.Sequence analysis of the complete genome of SDZQ-1 with that of twenty-one PCV-2 sequences published in GenBank indicated that the SDZQ-1 shows 95.0%-99.7%homology in complete nucleotide sequence compare with others.The two complete genome were connected into PUC19 subsequently in norientation to construct double copy plsmid pUC2PCV2.Transfect PK-15 cells with pUC2PCV2 in vitro.After four passages,the infectivity of the cloned chimeric PCV-2 genome was confirmed by PCR and IFA.

关 键 词：猪圆环病毒 感染性克隆 同源性 转染 

分 类 号：S852.65[农业科学—基础兽医学]