猪内源性逆转录病毒在3种不同品系猪中的分布及序列特征研究  

Distribution and sequence characteristics of porcine endogenous retroviruses in three different breeds of pigs

作　　者：冯天悦 Kamal Shah 张文立 付利芝 李昌文[2] 王玉娥[2] 陈洪岩[2] FENG Tian-yue;Kamal Shah;ZHANG Wen-li;FU Li-zhi;LI Chang-wen;WANG Yu-e;CHEN Hong-yan(School of Life Sciences,Northeast Agricultural University,Harbin 150030,China;State Key Laboratory of Veterinary Biotechnology,Harbin Veterinary Research Institute,Chinese Academy of Agricultural Sciences/Innovative Team for Laboratory Animals and Comparative Medicine/National Poultry Laboratory Animal Resource Bank,Harbin 150069,China;Institute of Veterinary Sciences&Pharmaceuticals,Chongqing Academy of Animal Sciences,Chongqing 402460,China)

机构地区：[1]东北农业大学生命科学学院,黑龙江哈尔滨150030 [2]中国农业科学院哈尔滨兽医研究所,兽医生物技术国家重点实验室/黑龙江省实验动物与比较医学重点实验室/国家禽类实验动物资源库,黑龙江哈尔滨150069 [3]重庆市畜牧科学研究院兽医兽药研究所,重庆402460

出　　处：《中国预防兽医学报》2022年第11期1141-1148,1168,共9页Chinese Journal of Preventive Veterinary Medicine

基　　金：黑龙江省重点研发计划指导类项目(GZ20210010)。

摘　　要：为研究猪内源性逆转录病毒(PERV)在不同猪种中的分布情况、序列结构、遗传变异、表达特性等特征,本研究选取大白猪、荣昌猪、五指山小型猪3种常见猪种为研究对象,分别设计特异性引物,检测各猪种肺组织中PERV 3种亚型(PERV-A、PERV-B、PERV-C)的分布情况。结果显示,PERV在3种猪种中均有分布,其中PERV-A分布最广泛。进一步分段扩增3种猪的PERV-A全基因组序列并测序分析。全基因组序列进化树与囊膜(env)基因进化树分析均显示,东北地区的大白猪与荣昌猪PERV-A归为同一分支,亲缘关系较近;对测序结果进行生物信息学分析和蛋白二级结构预测,结果显示,PERV-A Gag蛋白和Pol蛋白的变异程度较大,有多种结构,但Env蛋白保守性较高,结构较为单一。采用荧光定量PCR法检测SPF大白猪不同组织中的PERV-A,结果显示从SPF大白猪心、肝、脾、肺、肾、扁桃体、淋巴结等7种组织中均检测出PERV-A,且在扁桃体等免疫器官组织中的转录水平最高。本研究共获得了5条不同猪种的PERV-A全基因组序列,首次鉴定了荣昌猪PERV全基因组序列,提示东北地区大白猪的引种来源可能与韩国猪种存在一定同源性,且五指山小型猪与荣昌猪、大白猪亲缘关系较远,有助于PERV分子特性的研究,并为评估不同猪种PERV在异种移植中的安全性并筛选合适猪种提供参考。To investigate the distribution,sequence structure,genetic variation and expression characteristics of porcine endogenous retrovirus(PERV) in different pig breeds,this study selected three common pig breeds,namely the large white,Rongchang and Wuzhishan miniature pigs,as the study materials,and designed specific primers to detect the presence of three subtypes of PERV.The results showed that PERV is distributed in all three pig breeds,and the PERV-A is the most widely distributed subtype.The full sequences of the PERV-A isoforms from the three pig species were further amplified and sequenced.Both of the full sequence evolutionary tree and the envelope(env) gene evolutionary tree analyses showed that there is a close kinship between the large white and Rongchang pigs PERV-A subtype.The bioinformatics analysis and protein secondary structure prediction results showed that the Gag and Pol proteins of PERV-A subtype are highly variable and have multiple structures,but the Env protein is more conserved and have single structure.The expression of PERV-A isoforms in different tissues of SPF large white pigs was detected by fluorescence quantitative PCR.The results showed that PERV-A isoforms is present in six tissues of SPF pigs,including heart,liver,spleen,lung,kidney,tonsils and lymph nodes,and the highest expression is in immune organ tissues such as tonsils.In this study,a total of five full sequences of PERV-A subtypes from different pig breeds were obtained,and for the first time the full gene sequence of PERV from Rongchang pigs were identified.Besides,the origin of the large white pigs in Northeast China was postulated,and the genetic characteristics of small pigs in Wuzhishan indicated that these pigs had a far kinship with large white and Rongchang pigs.Our data contribute to the study of the molecular characteristics of PERV and provide a reference for assessing the safety of xenotransplantation due to PERV in different pig breeds and screening for suitable pig breeds.

关 键 词：猪内源性逆转录病毒 序列分析 囊膜基因 遗传进化分析 

分 类 号：S852.65[农业科学—基础兽医学]