期刊论文

Construction of infectious clone of ORF3 deleted strain of porcine circovirus type2a

中文

中国预防兽医学报

1008-0589

2017

39

5

351-355

10.3969/j.issn.1008-0425.2017.05.04

国家自然科学基金项目（31372406） 湖南省研究生科研创新项目（CX2016B285）

本校为第一机构

为构建猪圆环病毒Ⅱ型(PCV2)的感染性克隆及其ORF3缺失的感染性克隆,本研究通过比较分析GenBank中8种亚型的PCV2基因组序列(各3株),设计了PCV2基因全长克隆引物,以从江西省萍乡市某猪场疑似感染断奶仔猪多系统衰竭综合征病死猪的病料中提取的DNA为模板,利用PCR方法扩增PCV2全长基因序列并进行测序。利用分子生物学软件对测序结果分析,结果表明所获得的克隆为PCV2a-2A基因型。采用重叠延伸PCR(SOE-PCR)方法扩增病毒全长基因组DNA,克隆于pSP72载体中构建野生型PCV2a病毒株感染性克隆。此外,采用SOE-PCR方法,缺失野生型PCV2a感染性克隆中的阅读框3 (ORF3),并将这两种重组质粒分别转染PK15细胞,盲传...

To construct the infectious clone of porcine circovirus type 2 (PCV2), the total DNAs were extracted from pig carcasses samples in the field from Pingxiang region of Jiangxi province, which were suspected to die of post weaning multisystemic wasting syndrome (PMWS). Then, PCV2 genomic DNA was amplified from the sample by PCR and sequencing. DNA sequence and homology analysis suggest the PCV2 was classified into PCV2a-2A sub-genotype. Based on the viral sequences, two infectious PCV2 DNA clones were constructed, one of which was the wild type of PCV2a-2A and another one was the same as the wild...