摘要
从贵州三穗县某鸭场的病鸭病料中分离出1株坦布苏病毒(tembusu virus,TMUV),经鉴定将其命名为TMUV-GZ。为了解该分离株的生物学特性及其E基因的分子特点,测定TMUV-GZ株的半数胚致死量(ELD50),将病毒接种至不同宿主细胞,观察其适应性与增殖能力;对E基因克隆测序并进行序列分析,对其编码的蛋白质进行生物信息学分析。结果表明:TMUV-GZ株能导致鸡胚和鸭胚死亡,该分离株无血凝性,在鸭胚上的ELD50为10^(-2.63)·(0.2 mL)^(-1);该分离株能在Vero细胞、BHK-21细胞和DF-1细胞上增殖,对BHK-21细胞适应性最好。该分离株的E基因与泰国分离株DK/TH/CU-99核苷酸同源性最高,为98.3%;一级结构预测E蛋白的理论等电点为5.01,不稳定系数为39.5,为稳定蛋白质;二级结构预测该蛋白质以α-螺旋、β-折叠、β-转角较多,为亲水性蛋白质,可能存在10个优势抗原表位、2个跨膜区、3个结构域,无信号肽;三级结构预测该蛋白质模型结构与其他黄病毒E蛋白实验结构之间具有良好的一致性。综上,这一研究为TMUV亚单位疫苗和基因工程疫苗的研制提供了理论依据。
A strain of tembusu virus named TMUV-GZ was isolated from the diseased duck samples from a duck farm in Sansui County,Guizhou Province in this study.In order to understand the biological characteristics of the isolate and the molecular characteristics of the E gene,the ELD_(50)of the TMUV-GZ strain was determined in this study,and the virus was inoculated into different host cells to observe its adaptability and proliferation ability.The E gene was cloned and sequenced,and bioinformatics methods were used for sequence analysis,and the encoded protein was analyzed by bioinformatics analysis.The results showed that the TMUV-GZ strain can cause the death of chicken embryos and duck embryos,had no hemagglutination and the ELD_(50)was 10~(-2.63)·(0.2mL)^(-1)on duck embryos.The isolate can proliferate on Vero cells,BHK-21 cells and DF-1 cells,and had the best adaptability to BHK-21 cells.The nucleotide of the E gene of the isolate had the highest homology with the Thai strain DK/TH/CU-99,which was 98.3%.The primary structure predicted that the theoretical isoelectric point and instability coefficient of the E protein were 5.01 and 39.5 respectively,which indicated that it was a stable protein.The secondary structure predicted that the protein was hydrophilic protein,with moreα-helix,β-sheet,andβ-turn,and there may be 10 dominant epitopes,2 transmembrane regions,3 structural domains and no signal peptide.The tertiary structure predicted that this protein model was in good agreement with the experimental structure of other flavivirus E proteins.The results of this experiment would provide a theoretical basis for the development of TMUV subunit vaccine and genetic engineering vaccine.
作者
徐景峨
胡成云
陈汝佳
李婷
张亚楠
余波
张雨梅
李林林
XU Jing'e;HU Chengyun;CHEN Rujia;LI Ting;ZHANG Ya'nan;YU Bo;ZHANG Yumei;LI Linlin(College of Veterinary Medicine,Yangzhou University,Yangzhou 225009,China;Institute of Animal Husbandry and Veterinary Science,Guizhou Academy of Agricultural Sciences,Guiyang 550005,China;Yangzhou Yangda Lianhuan Pharmaceutical Genetic Engineering Company Limited,Yangzhou 225001,China;College of Animal Science,Guizhou University,Guiyang 550005,China;Collaborative Innovation Center for Prevention and Control of Important Animal Epidemics and Zoonoses in Jiangsu Province,Yangzhou 225009,China;Institute of Animal Health,Guangdong Academy of Agricultural Science,Guangdong 510640,China)
出处
《扬州大学学报（农业与生命科学版）》
CAS
北大核心
2023年第2期10-18,共9页
Journal of Yangzhou University：Agricultural and Life Science Edition
基金
贵州省科技计划项目[黔科合支撑(2022)一般005]
贵州省科技计划项目[黔科合支撑(2019)2287号]
江苏高校优势学科建设工程项目(PAPD)
国家现代农业产业技术体系专项(CARS-42-53)。
