Ⅰ型蓝舌病毒NS4基因的克隆及序列分析  

Cloning and Sequence Analysis ofⅠSerotype Bluetongue Virus NS4 Gene

作　　者：赵瑶 徐栋 马三立 马鲜平 王芝英 张德志[1,2] 李前勇 谢成华[3] 杨发辉 易华山 ZHAO Yao;XU Dong;MA San-li;MA Xian-ping;WANG Zhi-ying;ZHANG De-zhi;LI Qian-yong;XIE Cheng-hua;YANG Fa-hui;YI Hua-shan(College of Veterinary Medicine,Southwest University,Rongchang Chongqing 402460,China;Chongqing Veterinary Science Engineering Research Center,Rongchang Chongqing 402460,China;Chongqing Chengkou Vocational Education Center,Chengkou Chongqing 405900,China;Chongqing Rongchang Agricultural Technical Extension Station,Rongchang Chongqing 402460,China)

机构地区：[1]西南大学动物医学院,重庆荣昌402460 [2]重庆市兽医科学工程研究中心,重庆荣昌402460 [3]重庆市城口县职业教育中心,重庆城口405900 [4]重庆市荣昌区农业技术推广站,重庆荣昌402460

出　　处：《西南大学学报（自然科学版）》2020年第9期77-86,共10页Journal of Southwest University(Natural Science Edition)

基　　金：中央高校基本科研业务专项基金项目(XDJK2018C059,XDJK2018C060);重庆市基础研究与前沿探索专项基金项目(cstc2018jcyjAX0615).

摘　　要：为研究蓝舌病毒NS4基因分子特征,本研究从重庆分离的蓝舌病毒(BTV-1/China/2017)感染细胞提取总RNA,采用RT-PCR方法对蓝舌病毒(BTV-1/China/2017)NS4基因进行克隆与序列分析.结果表明:所获得的NS4基因ORF长234 bp,编码77个氨基酸.通过DNAMAN软件分析,表明本研究中克隆的NS4基因与BTV-8/Netherland/2008/03,BTV-4/Italy/2003,BTV-4/Israel/2010,BTV-8/Israel/2010/03,BTV-1/France/2007/18,BTV-1/France/2008/24,BTV-1/Algeria/2006/01,BTV-2/Sudan/1983/05,BTV-14/Cameroon/1982/04株相似性为100%,与BTV-2/South Africa/RSA vaccine strain,BTV-9/South Africa/RSA vaccine strain株及BTV-1/Libya/2007,BTV-2/Italy/2000等25株参考毒株的相似性为98.7%,而与BTV-1/Australia/2003/01和BTV-16/Indonesia/1998相似性最低,为90.8%;突变主要发生在5~6位、17~18位、35~36位、39~40位、53~54位、65位及73~74位.系统进化分析表明:BTV-1/China/2017株NS4基因与欧洲源BTV-4/Italy/2003,BTV-8/Netherland/2008/03,BTV-2/France/2001,BTV-15/Israel/2006,BTV-4/Israel/2010,BTV-1/France/2008/24,BTV-4/Spain/2005株聚为一类,同BTV-4/Italy/2003株亲源关系最近.通过在线软件预测发现NS4蛋白存在2个α-螺旋区,无跨膜区,具有亮氨酸拉链(bZIP)典型结构;存在典型的核定位信号并具有线粒体亚细胞定位特征.In order to understand the molecular characteristics of the NS4 gene of bluetonge virus(BTV),its total RNA was extracted from the cells infected by the BTV isolated from Chongqing(BTV-1/China/2017),and the NS4 gene of this virus strain was amplified with RT-PCR.The results showed that the amplified BTV NS4 gene contained an ORF 234 bp in length and encoded 77 amino acids.Analysis with DNAMAN showed that the cloned NS4 gene shared a 100%similarity with the strains BTV-8/Netherland/2008/03,BTV-4/Italy/2003,BTV-4/Israel/2010,BTV-8/Israel/2010/03,BTV-1/France/2007/18,BTV-1/France/2008/24,BTV-1/Algeria/2006/01,BTV-2/Sudan/1983/05 and BTV-14/Cameroon/1982/04,a 98.7%similarity with BTV-2/South Africa/RSA vaccine strain,BTV-9/South Africa/RSA vaccine strain and BTV-1/Libya/2007 and BTV-2/Italy/2000 and other 21 reference strains,and a 90.8%similarity with the BTV-1/Australia/2003/01 strain and the BTV-16/Indonesia/1998 strain.The amino acid mutation sites were mainly distributed in 5-6,17-18,35-36,39-40,53-54,65 and 73-74.Phylogenetic analysis indicated that the NS4 gene of the cloned gene and BTV-4/Italy/2003,BTV-8/Netherland/2008/03,BTV-2/France/2001,BTV-15/Israel/2006,BTV-4/Israel/2010,BTV-1/France/2008/24 and BTV-4/Spain/2005 strain isolated from European countries were located in the same evolutionary branch,which showed that they had a close phylogenetic relationship with BTV-4/Italy/2003 strain.Prediction of NS4 proteins through online software identified two alpha helixes and a typical leucine zipper(bZIP)structure,but no transmembrane region.They had a typical nuclear localization signal and the character of mitochondrial subcellular localization.

关 键 词：蓝舌病毒 NS4基因 序列分析 

分 类 号：S852.65[农业科学—基础兽医学]