山羊内源性鼻内肿瘤病毒enENTV-FJ1株全基因组测序及生物信息学分析  

Whole genome sequencing and bioinformatics analysis of Endogenous enzootic nasal tumor virus strain enENTV-2FJ1

作　　者：江锦秀[1] 张靖鹏 林裕胜 毛坤明[2] 游伟[1] 黄丽丽[3] 胡奇林[1] JIANG Jin-xiu;ZHANG Jin-peng;LIN Yu-sheng;MAO Kun-ming;YOU Wei;HUANG Li-li;HU Qi-lin(Institute of Animal Husbandry&Veterinary Medicine,Fujian Academy of Agricultural Sciences,Fuzhou 350013,China;Fuqing Technical Service Center of Animal Husbandry and Veterinary Medicine,Fuzhou 350300,China;Fujian Normal University,Fuzhou 350108,China)

机构地区：[1]福建省农业科学院畜牧兽医研究所,福建福州350013 [2]福清市畜牧兽医技术服务中心,福建福州350300 [3]福建师范大学,福建福州350108

出　　处：《中国预防兽医学报》2020年第10期1062-1067,共6页Chinese Journal of Preventive Veterinary Medicine

基　　金：福建省科技计划项目-省属公益类科研院所重点项目(2018R1023-1);家畜疫病快速诊断技术及新疫苗研发与应用(2019NZ09007);福建省农业科学院科技创新团队建设项目(STIT2017-1-5)。

摘　　要：为了丰富山羊内源性鼻内肿瘤病毒(enENTV)全基因组序列资源,本研究从1份患羊地方性鼻内肿瘤(ENT)病羊的肾脏中经PCR扩增了1株enENTV全基因组序列,命名为enENTV-FJ1,分别对其全基因组、5’端长末端重复序列(5’LTR)基因、gag基因、env基因及其编码的氨基酸序列进行同源性分析并构建系统进化树,对enENTV的5’LTR进行酶切位点分析,并分析该株病毒的gag、env序列与enENTV、山羊地方性鼻内肿瘤病毒(ENTV-2)、绵羊鼻内肿瘤病毒1型(ENTV-1)等相关病毒参考株相应序列的差异性。结果显示,enENTV-FJ1基因组结构为LTR-gag-pro-pol-env-LTR,长度为7923 bp;全基因组同源性分析结果显示,enENTV-FJ1株与enENTVCHN1~enENTV-CHN66个参考株的同源性为95.7%~97.3%,同源性最近;与ENTV-2参考株的同源性为87.0%~93.2%;enENTV-FJ1的5’LTR基因序列、gag基因及其编码的氨基酸序列与enENTV参考株相应序列的同源性分别为92.8%~99.1%、92.6%~97.5%、92.2%~97.7%;enENTV-FJ1 env基因及其编码的氨基酸序列与内源性绵羊肺腺瘤病毒(enJSRV)参考株相应序列的同源性分别为93.3%~93.8%、94.3%~94.8%。全基因组进化树结果显示,enENTV-FJ1与enENTV-CHN1~enENTV-CHN6处于同一个进化分支;5’LTR进化树结果显示,enENTV-FJ1株与enENTV-CHN2处于同一进化分支;gag基因进化树结果显示enENTV-FJ1与ENTV-CHN9处于一个小的进化分支,与enENTV-CHN6同处于一个较大的进化分支;env基因进化树结果显示,enENTV-FJ1株与ENTV-2/Spain株处于同一进化分支;所有enENTV的5’LTR区均无ENTV-2参考株所含有的Hpy188Ⅰ、NlaⅣ、MowⅠ这3个酶切位点。gag基因序列分析结果显示,与enJSRV、外源ENTV-2、ENTV-1及绵羊肺腺瘤病毒(JSRV)的gag基因相比,enENTV-FJ1株在697 bp^705 bp有9个核苷酸的插入,该基因编码的氨基酸在aa229~aa232处插入3个脯氨酸。env基因编码的氨基酸结果显示,enENTV-FJ1株与ENTV-2株在aa1~aa7处插入了7个氨基酸(MFVFFRR)。�To enrich the resource of the whole genome sequence of Endogenousenzootic nasal tumor virus(enENTV),the whole genome sequence of one strain of enENTV was amplified by PCR in the kidney sample of a goat suffering from enzootic nasal tumor(ENT)disease and named enENTV-FJ1 in this study.The nucleotide and amino acid sequence homology analysis of the whole-genome,5’LTR gene,gag and env genes were performed to construct a phylogenetic tree,respectively,and the restriction enzyme site analysis was conducted on the 5’LTR gene of enENTV and ENTV-2.The differences between the gag,env and the corresponding sequence of the related virus reference strains such as enENTV,ENTV-2,ENTV-1,enJSRV and JSRV were analyzed,respectively.The results showed that the genome of enENTV-FJ1 were 7923 base pairs(bp)in length with the LTR-gag-pro-pol-env-LTR genomic structure,which shared 95.7%-97.3%identity of nucleotide sequences with the six strains of enENT-VCHN1 enENTV-CHN6 and 87.0%-93.2%identity of nucleotide sequences with ENTV-2 reference strain.The enENTV-FJ15’LTR gene shared 92.8%-99.1%identity with that of the enENTV reference strain,gag gene and its encoded amino acid sequence of enENTV-FJ1 shared 92.6%-97.5%and 92.2%-97.7%identity of the corresponding sequence of the enENTV reference strain,respectively.The enENTV-FJ1 env gene and its encoded amino acid shared 93.3%-93.8%and 94.3%-94.8 identity of the corresponding sequence of the enJSRV reference strain,respectively.The gene phylogenetic analysis showed that enENTV-FJ1 and enENTV-CHN1 enENTV-CHN6 clustered into the same branch,the 5’LTR gene of the enENTV-FJ1 virus strain and enENTV-CHN2 belonged to the same branch,the gag gene of enENTV-FJ1 and ENTV-2 CHN9 clustered into a small branch,and enENTV-CHN6 clustered into the large branch.The env gene of enENTV-FJ1 and ENTV-2/Spain clustered the same branch.The 5’LTR region of enENTV-FJ1 didn’t possess the three restriction sites,Hpy188Ⅰ,NlaⅣ,and MowⅠ,but ENTV-2 virus strain did.The gag gene sequence analysis result

关 键 词：山羊 内源性山羊鼻内肿瘤病毒 全基因组测序 生物信息学分析 

分 类 号：S855.3[农业科学—临床兽医学]