摘要
旨在对鸭腺病毒3型(duck adenovirus 3,DAdV-3)广东流行株进行分离,并预测分析六邻体(Hexon),五邻体(Penton),纤维蛋白(Fiber1、Fiber2)4个主要结构蛋白的理化性质、结构及功能。本研究用鸡肝癌细胞(LMH)对疑似DAdV-3感染的鸭、鹅肝脏组织进行病毒分离,对3株病毒分离株主要结构蛋白进行基因克隆、测序、拼接,获得4个蛋白的核苷酸序列后,使用MEGA 11软件进行氨基酸序列分析并构建遗传进化树,利用Expasy-ProtParam与SOPMA软件分析4个蛋白的理化性质与二级结构,应用AlphaFold2软件预测4个蛋白的三级结构,并对4个蛋白的线性B细胞抗原表位进行预测。结果:成功分离得到3株DAdV-3,命名为XZD-2021株、FRK-2021株、825-1-2021株;3株DAdV-3的4个蛋白均与GenBank中其他DAdV-3分离株处于同一分支中;Hexon蛋白更接近鹅腺病毒4型,Penton蛋白更接近D种禽腺病毒,Fiber1、Fiber2蛋白则更接近鸽腺病毒1型;4个蛋白均为酸性亲水蛋白,无信号肽,Hexon、Fiber1、Fiber2蛋白以自由卷曲为主,Penton蛋白则以α螺旋为主;三级结构预测结果与二级结构结果相符,线性B细胞抗原表位预测发现,Hexon蛋白的线性B细胞抗原表位最多,为11个,Penton蛋白与Fiber2蛋白为6个,Fiber1蛋白为5个,但Fiber1蛋白位于病毒表面的抗原表位最多,表位长度较Fiber2更长,且多位于负责与宿主细胞受体结合的头节区。综上,本研究成功分离得到3株DAdV-3,并对其4个主要结构蛋白进行了遗传演化、二级结构、三级结构等生物学特性进行分析,研究结果为后续DAdV-3致病特性和免疫特性研究奠定基础。
This study aimed to isolate the Guangdong epidemic strain of duck adenovirus 3(DAdV-3),and to predict and analyze the physicochemical characteristics,structure,and function of its four major structural proteins(Hexon,Penton,Fiber1,and Fiber2).Suspected DAdV-3 infected duck and goose liver tissue samples were collected and subjected to virus isolation using chicken hepatoma cells(LMH).The major structural proteins(see above)of the four viral isolates were cloned,sequenced,and assembled.The nucleotide sequences of the four proteins were analyzed using the MEGA 11 software so as to perform amino acid sequence analysis and to construct phylogenetic trees.The physicochemical properties and secondary structures of the four proteins were analyzed using the Expasy-ProtParam and SOPMA software.The tertiary structures of the proteins were predicted using the AlphaFold2 software,and their linear B-cell epitopes were predicted.The results showed that three DAdV-3 virus strains were successfully isolated and were named XZD-2021,FRK-2021,and 825-1-2021,respectively.The four proteins of the three DAdV-3 strains were in the same branch as other DAdV-3 isolates in GenBank.The Hexon protein was closer to Goose adenovirus type 4,the Penton protein was closer to type D avian adenovirus,and the Fiber1 and Fiber2 proteins were closer to Pigeon adenovirus type 1.All four proteins were acidic hydrophilic proteins without signal peptides.The Hexon,Fiber1,and Fiber2 proteins were composed of random coils,while the Penton protein was predominantly ofα-helix.The tertiary structure predictions were consistent with the secondary structure results.The linear B-cell epitope prediction revealed that the Hexon protein had the most(11)linear B-cell epitopes,followed by the Penton and Fiber2 proteins(6 each),and the Fiber1 protein(5).However,Fiber1 had the most surface antigen epitopes with higher lengths than Fiber2,which was primarily located in the head domain responsible for binding to host cell receptors.In summary,in this study,three DAdV-3 strains were successfully isolated and the genetic evolution,secondary structure,and tertiary structure of their four major structural proteins were analyzed.The results provided a foundation for further research.
作者
陈作鑫
姜雪芹
刘梦凡
赵梓桥
魏无缺
李树稳
张雪莲
刘兆洁
黄淑坚
梅堃
CHEN Zuoxin;JIANG Xueqin;LIU Mengfan;ZHAO Ziqiao;WEI Wuque;LI Shuwen;ZHANG Xuelian;LIU Zhaojie;HUANG Shujian;MEI Kun(College of Life Science and Engineering,Foshan University,Foshan 528231,China;Guangdong Huasheng Biotechnology Co.,Ltd.,Guangzhou 510000,China)
出处
《畜牧与兽医》
CAS
北大核心
2024年第9期61-69,共9页
Animal Husbandry & Veterinary Medicine
基金
广东省重要新发水禽疫病防控共性关键技术集成与应用研究项目(2020ZDZX1004)
广东省水禽产业技术体系创新团队项目(2023KJ137)。
关键词
鸭腺病毒
分离鉴定
遗传进化分析
生物信息学
duck adenovirus
isolation and identification
genetic evolution analysis
bioinformatics
