河南省猪伪狂犬病病毒gC、gE基因组的克隆与遗传变异分析  

Cloning and phylogenetic variation analysis of the pseudorabies virus gC and gE genomes in Henan Province

作　　者：孔令昊 苗信永 赵士杰 李闻 徐朋丽 孙杨扬 崔志莹 夏平安[1] 张宜娜[1] 吴斌[2] KONG Linghao;MIAO Xinyong;ZHAO Shijie;LI Wen;XU Pengli;SUN Yangyang;CUI Zhiying;XIA Pingan;ZHANG Yina;WU Bin(College of Veterinary Medicine,Henan Agricultural University,Zhengzhou 450002,China;College of Veterinary Medicine,Huazhong Agricultural University,Wuhan 430070 China)

机构地区：[1]河南农业大学动物医学学院,河南郑州450002 [2]华中农业大学动物医学学院,湖北武汉430070

出　　处：《中国兽医学报》2021年第5期858-864,共7页Chinese Journal of Veterinary Science

基　　金：“十三五”国家重点研发计划资助项目(2018YFD0500800)。

摘　　要：为了解近几年河南省猪伪狂犬病病毒(pseudorabies virus, PRV)的遗传进化情况,本研究对2018-2020年采自河南省部分地区的307份病料样品进行双重PCR检测,并进行gC、gE全基因组克隆和序列分析。结果显示307份样品中有14份呈阳性,其阳性率为4.56%,且从14份阳性样品中分别克隆了gC、gE全基因组。核苷酸序列分析结果显示,gC基因之间的核苷酸同源性为99.6%~100.0%,与国内变异毒株HeN1-2012等的核苷酸同源性为99.8%~100.0%,与国外经典毒株Bartha等的核苷酸同源性为94.7%~95.8%;gE基因之间核苷酸同源性为98.7%~100%,与国内变异毒株HeN1-2012等的核苷酸同源性为98.7%~99.9%,与国外经典毒株Bartha等的核苷酸同源性为96.6%~97.7%。氨基酸序列对比结果显示,本研究测序14株PRV与国内经典毒株Ea株相比,gC基因有3处氨基酸突变(N34T、K99E、E194G),gE基因存在自6处氨基酸突变(G54D、P403A、V450I、496D(插入)、G510S、S518P);与国内变异毒株HeN1-2012相比,均没有发生氨基酸位点突变;与国外经典毒株Bartha等经典毒株相比gC全基因均存在第63位点处连续插入7个氨基酸(AAASTPA),并且存在30处氨基酸的替换,gE全基因存在23处氨基酸变异位点。氨基酸序列分析显示本研究测序14株PRV与国内变异毒株HeN1-2012处于同一分支,亲缘关系最近,且这些特征性变异位点可作为分子诊断依据,为河南省猪场PRV的净化提供了参考依据。In order to understand the genetic evolution of porcine pseudorabies virus(PRV)in Henan province in recent years, this study conducted a double PCR test on 307 disease samples collected from parts of Henan province from 2018 to 2020,and gC and gE whole genome cloning and sequence analysis were performed.The results showed that 14 of the 307 samples were positive, the positive rate was 4.56%,and gC and gE whole genomes were cloned from 14 positive samples.The nucleotide sequence analysis results show that the nucleotide homology between gC genes was 99.6%-100.0%,and the nucleotide homology with domestic mutant strains such as HeN1-2012 was 99.8%-100.0%,which was similar to foreign classic strains.The nucleotide homology of Bartha et al.is 94.7%-95.8%;the nucleotide homology between gE genes is 98.7%-100.0%,and the nucleotide homology with domestic mutant strains such as HeN1-2012 is 98.7%-99.9%,the nucleotide homology with the classical foreign strain Bartha etc.is 96.6%-97.7%.Amino acid sequence comparison results showed that compared with the domestic classic strain Ea strain,14 PRVs sequenced in this experiment had 3 amino acid mutations in the gC gene(N34 T,K99 E,E194 G),and there were 6 amino acid mutations in the gE gene(G54 D,P403 A,V450 I,496 D(insertion),G510 S,S518 P);compared with the domestic mutant strain HeN1-2012,no amino acid site mutations occurred;compared with the foreign classic strain Bartha,there were 7 consecutive amino acids(AAASTPA)inserted at position 63 of the gC gene,and there were 30 amino acid substitutions.There were 23 amino acid mutation sites in the gE gene.Amino acid sequence analysis showed that 14 PRVs sequenced in this experiment were in the same branch as the domestic mutant strain HeN1-2012,and they had the closest genetic relationship.These characteristic mutation sites can be used as a molecular diagnosis basis,which provides a reference for the elimination of PRV in pig farms in Henan.

关 键 词：猪伪狂犬病毒 序列分析 GC gE 

分 类 号：S852.65[农业科学—基础兽医学]