PCV-2 GZ-BJJS2015株感染性克隆质粒的构建  

Construction of infectious cloning plasmid of PCV-2 GZ-BJJS2015 strain

作　　者：咸文 梁海英[1] 曾智勇[1] 王彬[1] 刘钊[1] 徐国[1] 叶百川 张爱琼[1] 何小莉[1] XIAN Wen;LIANG Haiying;ZENG Zhiyong;WANG Bin;LIU Zhao;YE Baichuan;ZHANG Aiqiong;HE Xiaoli(College of Animal Science, Guizhou University, Guiyang 550025, Chin)

机构地区：[1]贵州大学动物科学学院

出　　处：《黑龙江畜牧兽医》2018年第11期16-19,共4页Heilongjiang Animal Science And veterinary Medicine

基　　金：国家自然科学基金项目(31560688)

摘　　要：为了解贵州地区猪圆环病毒2型（PCV-2）的遗传变异情况,丰富贵州省PCV-2的病毒库,试验应用PCR反向遗传操作技术构建感染性克隆质粒,从送检的病毒血症样品中拯救获得1株PCV-2毒株,将其命名为PCV-2 GZ-BJJS2015,并进行测序及比对分析。结果表明：该病毒基因组大小为1 767 bp,属于PCV-2b型;与参考株pmws（AF027217）相比,其ORF5在1 042位缺失1个碱基T,ORF9在第92位发生基因突变,致使ORF9不能编码蛋白;ORF5与病毒毒力有一定关系,其变异可能会影响毒株毒力,ORF9不编码蛋白的意义有待考究。根据强弱毒株特征性氨基酸位点的差异比较结果,可初步推测该毒株为PCV-2强毒株。The aim of the present study was to understand the genetic variation of Porcine circovirus type 2（ PCV-2） in Guizhou area and enrich PCV-2 virus library of the province. The infectious cloning plasmid was constructed by using the PCV reverse genetic manipulation technology.A PCV-2 strain was obtained from the viremia sample,named PCV-2 GZ-BJJJS2015,and sequenced for comparison analysis. The results showed that the genome size of the rescued virus was 1 767 bp,which belonged to PCV-2b type. Compared with the reference strain pmws（ AF027217）,the ORF5 lacked one base T at 1 042. Gene mutation occurred in ORF9 at the 92 nd position of the gene sequence,resulting in ORF9 being unable to encode protein. ORF5 was related to virulence of PCV-2,and its variation might affect virulence of strain. Further studies were needed to confirm the significance of ORF9 gene silent. According to the difference of the characteristic amino acid sites of the strong and weak strain,the strain was speculated to be a PCV-2 virulent strain.

关 键 词：猪圆环病毒2型 反向遗传操作技术 病毒拯救 感染性克隆质粒 分离鉴定 

分 类 号：S852.659.2[农业科学—基础兽医学]