5株PCV2四川株全基因组的克隆与遗传进化分析  

Cloning and phylogenetic analysis of complete genome of five PCV2 strains from Sichuan

作　　者：杨泽晓[1,2] 马玉峰[1,3] 汪开毓[1] 王印[1,2] 赵希仑 姚学萍[1] 任冉阳[1] 陈平[1] 胡凌[1] 

机构地区：[1]四川农业大学动物医学院,四川温江611130 [2]动物疫病与人类健康四川省重点实验室,四川温江611130 [3]金宇保灵生物药品有限公司,内蒙古呼和浩特010030

出　　处：《西北农林科技大学学报（自然科学版）》2016年第4期1-9,16,共10页Journal of Northwest A&F University(Natural Science Edition)

基　　金："十二五"国家科技支撑计划项目(2013BAD12B04);四川农业大学双支计划项目(00270401)

摘　　要：[目的] 了解目前四川省猪圆环病毒2型（PCV2）的分子流行病学特征和遗传变异情况,探讨PCV2感染的控制对策。[方法] 根据GenBank中发表的PCV2基因组序列设计2对特异性引物,对2012-2014年临床送检的PCV2感染病例材料中的5株PCV2全基因组序列进行扩增测序,并利用DNAStar等生物信息分析软件对获得的PCV2基因组序列进行分子特性与遗传演变分析。[结果] 5株PCV2四川株全基因组序列长度都为1 767bp,均具有滚环复制起点（ORC）典型结构。序列比对和进化分析显示,5株PCV2四川株的基因序列同源性很高,基因组核苷酸序列及ORF1、ORF2和ORF3基因序列同源性分别为98.4%～99.9%,97.5%～99.6%,99.4%～100%和96.2%～100%,ORF1、ORF2和ORF3基因推导氨基酸序列同源性分别为98.4%～99%,99.6%和91.5%～100%;与47条参比PCV2毒株核苷酸序列的同源性比较发现,5株PCV2与国内一些新出现的PCV2毒株亲缘关系较近,且均属于PCV2d基因型毒株。[结论] 5株PCV2四川株之间尽管存在着不同程度的基因变异,但遗传进化相对较稳定。[Objective] This study investigated the molecular epidemiology characteristics and genetic heterogeneity of PCV2 in Sichuan to provide reference for control and prevention of PCV2 infection. [Method] Two pairs of special primers were designed and synthesized according to the full genome se- quences of PCV2 published in GenBank. Then they were used to amplify and sequence the full genome se- quences of 5 PCV2 strains in clinical samples of PCV2 infection cases collected from Sichuan during 2012 to 2014 using PCR. Molecular epidemiology characteristics and genetic heterogeneity of 5 PCV2 were also analyzed using DNAStar and MEGA 4.1. [Result] The genomic sequences of 5 PCV2 strains were all 1 767 bp in the length and had the classic ORC structure in sequence. Phylogenetic analysis showed that the ge- nomes of 5 PCV2 Sichuan strains were closely related to each other. The nucleotide homology of PCV2 full length genome,ORF1, ORF2 and ORF3 were 98. 4% - 99. 9%, 97. 5% - 99. 6%, 99. 4% - 100% and 96.2%-100%, respectively. The amino acid sequence homologies of ORF1, ORF2 and ORF3 98.4%-99% ,99.6% and 91.5%-100% ,respectively. Phylogenetic analysis also showed that the 5 Sichuan strains were close to some recent PCV2 isolates in China and belonged to genotype PCV2d. [Conclusion] The evolution of PCV2 was stable although there were nucleotide mutations at different degree among the 5 strains in Sichuan.

关 键 词：PCV2 基因克隆 遗传进化分析 

分 类 号：S852.659.2[农业科学—基础兽医学]