一株H1N1亚型猪流感病毒全基因克隆及遗传特性分析  

Cloning and Genetic Characteristics Analysis of H1N1 SIV Genome

作　　者：孟慧芝 孙涛 岳志芹 王文斌[2] 

机构地区：[1]山东出入境检验检疫技术中心,山东青岛266000 [2]山西农业大学,山西太谷030801

出　　处：《动物医学进展》2015年第6期48-53,共6页Progress In Veterinary Medicine

基　　金：国家科技支撑计划项目(2012BAK11B00)

摘　　要：为快速获知流感病毒的全基因组序列,及时得知其遗传演化特性。设计11对通用条形码引物,采用RT-PCR法扩增A／swine／Shandong／01／2009（H1N1）株的8个基因节段,进行克隆测序和遗传进化分析。核酸序列测定结果显示,全基因组序列完整无缺失,但是基因组内2个位点突变导致PB1-F2、NS-1蛋白表达不完整,HA切割位点处氨基酸序列PSIQSR/GLF,无多个连续的碱性氨基酸,符合非致病性切割位点的特征。从绘制的各个片段进化树分析,此毒株与11株近5年在人和猪中流行的H1N1相似度高,HA的同源性达98.1%~99.7%,NA同源性98.6%~99.8%,与2009年暴发的A型H1N1流行株A/California/07/2009（H1N1）8个片段进行比对,核酸同源性高达99.1%~99.6%。To obtain complete genome sequence and genetic evolution characteristics of influenza virus quickly and timely, 11 pairs of universal barcode primers were designed. All the eight genes of A／swine／ Shandong／01／2009 （H1N1） strain were amplified by RT-PCR method, and sequenced respectively. The complete genome sequenc analysis showed no deletion was observed in the isolate strain. But 2 loci muta- ted, PB1-F2 and NS-1 proteins were not expressed completely. The amino acid sequence of cleavage site of HA is PSIQSR/GLF and no multiple consecutive basic amino acid, suggesting that the isolate strain was accord with the characteristics of non pathogenicity. The analysis of phylogenetic tree drawn from each fragment showed that homology of HA was up to 98.1%-99.7% and NA was up to 98.6%-99.8% com- pared with the other 11 strains in the human and porcine epidemic influenza A viruses in near 5 years. Com- pared with the epidemic strain of A/California/07/2009 （H1N1）, homology was up to 99.1%-99.6%.

关 键 词：猪流感病毒 H1N1 全基因组测序 特异位点分析 

分 类 号：S852.659.5[农业科学—基础兽医学]