蓝狐细小病毒VP2基因与NS1基因的克隆与序列分析  

Cloning and sequence analysis of VP2 gene and NS1 gene of blue fox parvovirus

作　　者：刘海防[1] 胡传伟[2] 谢之景[1] 倪长鹏[2] 贾赟[2] 姜世金[1] 张兴晓[1] 朱艳丽[1] 徐丽秋[3] 高玉峰[4] 

机构地区：[1]山东农业大学动物科技学院,山东泰安271018 [2]辽宁出入境检验检疫局技术中心,辽宁大连116001 [3]沈阳农业大学动物科技学院,辽宁沈阳116001 [4]大连工业大学生物与食品工程学院,辽宁大连116034

出　　处：《中国兽医学报》2009年第8期978-981,1012,共5页Chinese Journal of Veterinary Science

基　　金：山东农业大学青年科技创新基金资助项目(23204);国家质检总局资助项目(2005IK047)

摘　　要：根据GenBank上发表的犬细小病毒（Canine parvovirus,CPV）、猫细小病毒（Feline parvovirus,FPV）核酸序列,分别设计扩增VP2基因与NS1基因的特异性引物,采用PCR技术扩增蓝狐细小病毒（Blue fox parvovirus,BF-PV）泰安分离株（BFPV-TA）的VP2基因和NS1基因,将PCR产物克隆入pMD18-T载体,进行测序分析。结果显示,BFPV-TA VP2基因含有1个ORF,全长1 755 bp,共编码584个氨基酸,第219位氨基酸发生I→V（219I→V）改变,300G→P,411E→A;BFPV-TA NS1基因含有1个ORF,全长2 007 bp,共编码668个氨基酸,8个氨基酸残基发生改变,43R→H,135H→Y,172K→R,179A→E,350D→N,409I→V,574I→V,616D→V。BFPV-TA VP2基因与CPV和FPV参照株的同源性在98.4%~99.4%,BFPV-TA NS1基因与CPV和FPV参照株的同源性为98.6%~99.1%。VP2基因系统发生分析,BFPV-TA与FPV的亲源关系较为密切,但NS1基因系统发生分析,BFPV-TA成单独的分支。Two pairs of primers were designed based on the VP2 gene and NS1 gene of the strains of canine parvovirus and feline parvovirus according to GenBank. Using blue fox parvovirus Tai＇an isolate（BFPV-TA）as template, the VP2 gene and NS1 gene were amplified with PCR,which were cloned into pMD18-T vector,sequenced and ana- lyzed. As a result,the ORF of the VP2 gene of BFPV-TA was 1 755 bp and encoded 584 amino acids. The VP2 protein possessed three substitutions,2191I→V,300G→P and 411E→A. The ORF of the NS1 gene of BFPV-TA was 2 007 bp and encoded 668 amino acids. The NS1 protein had eight substitutions, 43R→ H, 135 H→Y, 172K→R, 179A →E,350D→N,409I→V,574I→V,616D→V. The nucleotide homologies of VP2 genes between BFPV-TA and the CPV, FPV references are from 98.4 % to 99.4%, the identity of NS1 from 98.6 % to 99.1 %. VP2 gene phylogenetic analysis shoued that CDV-FOX-TA and the CDV wild strains had the same ancestor. BFPV-TA and FPV had the nearer inherit distance,but the clear cluster was observed based on NS1 gene phylogenetic analysis.

关 键 词：蓝狐细小病毒 VP2基因 NS1基因 序列分析 

分 类 号：S852.65[农业科学—基础兽医学]