J、K亚群禽白血病病毒的分离和囊膜蛋白(gp85)基因序列分析  

Isolation and Membrane Protein(gp85)Gene Sequence Analysis of Avian Leukosis Virus Subgroup J and K

作　　者：周钊灿 游广炬 杨金易[2,3] 苏晓娜 高丽 王永强 郑世军[1] ZHOU Zhaocan;YOU Guangju;YANG Jinyi;SU Xiaona;GAO Li;WANG Yongqiang;ZHENG Shijun(National Key Laboratory of Veterinary Public Health Safety,Key Laboratory of Animal Epidemiology of the Ministry of Agriculture and Rural Affairs,College of Veterinary Medicine,China Agricultural University,Beijing 100193,China;College of Food Science,South China Agricultural University,Guangzhou 510642,China;Yunfu Subcenter,Guangdong Laboratory for Lingnan Modern Agriculture,WENS Foodstuff Group Co.,Ltd.,Yunfu 527100,China)

机构地区：[1]中国农业大学动物医学院兽医公共卫生安全全国重点实验室农业部动物流行病学重点实验室,北京海淀100193 [2]华南农业大学食品学院,广东广州510642 [3]温氏食品集团股份有限公司岭南现代农业科学与技术广东省实验室云浮分中心,广东云浮527100

出　　处：《中国兽医杂志》2024年第4期1-10,共10页Chinese Journal of Veterinary Medicine

基　　金：国家重点研发计划(2022YFD1300100);国家蛋鸡产业技术体系(NYCYTX41)。

摘　　要：为了解现阶段我国禽白血病病毒(ALV)的流行特征和演化趋势,本试验于2021—2022年从国内某鸡场采集蛋清ALV衣壳蛋白(p27)抗原阳性的鸡血浆,接种鸡胚成纤维细胞(DF-1)分离病毒并进行亚群鉴定;采用聚合酶链式反应(PCR)技术扩增分离株的囊膜蛋白(gp85)基因片段,并对其进行测序,将基因序列与ALV不同亚群毒株进行比对和遗传进化分析;选取氨基酸变化差异较大的分离毒株进行间接免疫荧光鉴定。结果显示,共分离到7株ALV毒株(3株J亚群,4株K亚群),ALV-J分离毒株CAU4932、CAU4860和CAU2259的gp 85基因片段长度为918 bp,编码306个氨基酸,分离毒株之间gp 85基因核苷酸同源性为90.5%~95.2%,gp85蛋白氨基酸同源性为95.1%~99.3%;ALV-K分离毒株CAU7049、CAU5006、CAU7176和CAU7168的gp 85基因片段长度为1008 bp,编码336个氨基酸,分离毒株之间gp 85基因核苷酸同源性为89.4%~92.3%,gp85蛋白氨基酸同源性为94.0%~99.7%。gp85蛋白氨基酸序列同源性比对发现,分离株gp85蛋白氨基酸序列均会发生突变,但高变区hr1和hr2所占比例较多,证实了gp85的高变性。在gp85蛋白B细胞抗原表位中存在部分位点的氨基酸突变,可能导致gp85蛋白抗原性发生变化。间接免疫荧光鉴定结果显示,5株分离毒株均可在DF-1细胞内复制并存在可被抗体识别的p27抗原表位。结果表明,本试验分离的7株ALV毒株是国内ALV-J和ALV-K流行毒株的突变株,不仅丰富了ALV基因组库资源,且为后续研究ALV的遗传变异特征和流行趋势等提供参考依据。In order to understand the epidemiological characteristics and evolutionary trends of avian leukosis virus(ALV)in China,plasma samples positive for ALV capsid protein(p27)antigen in egg white were collected from a chicken farm in China from 2021 to 2022.The virus was isolated by inoculating Douglas foster-1(DF-1)and subgroup identification was conducted.The membrane protein(gp85)gene fragments of the isolated strains were amplified by polymerase chain reaction(PCR)technology and sequenced.The gene sequences were compared and analyzed for genetic evolution with different subgroups of ALV strains.Strains with significant amino acid variations were selected for indirect immunofluorescence identification.The results showed that a total of 7 strains of ALV were isolated(3 strains of J subgroup and 4 strains of K subgroup).The gp 85 gene fragments of the ALV-J isolates CAU4932,CAU4860,and CAU2259 were 918 bp in length,encoding 306 amino acids,with nucleotide homology of 90.5%-95.2%and amino acid homology of 95.1%-99.3%among the isolated strains.The gp 85 gene fragments of the ALV-K isolates CAU5006,CAU7176,and CAU7168 were 1008 bp in length,encoding 336 amino acids,with nucleotide homology of 89.4%-92.3%and amino acid homology of 94.0%-99.7%among the isolated strains.Amino acid sequence homology analysis of the gp85 protein revealed mutations in all isolated strains,with a higher proportion in the variable regions hr1 and hr2,confirming the high variability of gp85.Amino acid mutations were observed in some sites of the gp85 protein B cell antigenic epitopes,which may lead to changes in the antigenicity of the gp85 protein.Indirect immunofluorescence identification results showed that all five isolated strains could replicate in DF-1 cells and had p27 antigen epitopes recognized by antibodies.The results indicate that the 7 strains of ALV isolated in this study are mutant strains of circulating ALV-J and ALV-K in China,enriching the genetic database of ALV and providing reference for subsequent studies on genetic varia

关 键 词：禽白血病病毒 J亚群 K亚群 gp 85基因 序列比较 

分 类 号：S852.659.3[农业科学—基础兽医学]