1型鸭肝炎病毒VP1基因的序列分析及分段表达

发布时间：2017-12-28 09:44

本文关键词：1型鸭肝炎病毒VP1基因的序列分析及分段表达　出处：《内蒙古农业大学》2016年硕士论文　论文类型：学位论文

【摘要】：为了解我国1型鸭肝炎病毒的基因变异情况,本试验对6株分离自山东地区的鸭肝炎病毒株SD4、SD10、SD15、SD37、SD38、SD40病毒液接种鸭胚尿囊腔,收集尿囊液,鉴定冻存备用。对其VP1基因进行序列测定,用MEGA5软件与GenBank中已公布的14株鸭肝炎病毒VP1序列进行比较分析,比较核苷酸的同源性并绘制进化树。结果表明,SD4株VP1基因与10株1型鸭肝炎病毒VP1基因的同源在84.7%～91.0%之间,SD10株VP1基因与10株1型鸭肝炎病毒VP1基因的同源在86.5%～92.2%之间,SD15株VP1基因与10株1型鸭肝炎病毒VP1基因的同源在84.9%～94.0%之间,SD37株VP1基因与10株1型鸭肝炎病毒VP1基因的同源在84.8%～90.3%之间,SD38株VP1基因与10株DHAV-1的同源性在92.2%-98.0%之间,SD40与10株1型鸭肝炎病毒VP1基因的同源性在85.9%～91.8%之间。6株试验毒株与2型和3型鸭肝炎病毒VP1基因的同源性在62.5%～71.6%之间,同源性较低。SD38 VP1基因与10株1型鸭肝炎病毒VP1基因的同源性较高。同时,本试验对SD38病毒株VP1基因进行分段表达。成功获得了高抗原性的可溶性蛋白pCold-TF-1Q、pCold-TF-1Z、pCold-TF-1H,并对获得的分段表达蛋白进行了纯化。本试验为建立检测Ⅰ型鸭病毒性肝炎抗体的ELISA检测方法奠定了基础。
[Abstract]:To understand the genetic variation of duck hepatitis virus type 1 in China, 6 strains of duck hepatitis virus SD4 isolated from Shandong, SD10, SD15, SD37, SD38 and SD40 were inoculated into the duck embryo allantoic cavity, and the allantoic fluid was collected for identification of frozen storage. The sequence of VP1 gene was sequenced. Compared with 14 published duck hepatitis virus VP1 sequences published in GenBank, MEGA5 was used to compare the nucleotide homology and draw the phylogenetic tree. The results showed that SD4 strain VP1 gene and 10 strains of duck hepatitis virus type 1 VP1 gene homology in 84.7% ~ 91% between SD10 strain VP1 gene and 10 strains of duck hepatitis virus type 1 VP1 gene homology in 86.5% ~ 92.2% between SD15 strain VP1 gene and 10 strains of duck hepatitis virus type 1 VP1 gene homology 84.9% ~ 94%, SD37 strain VP1 gene and 10 strains of duck hepatitis virus type 1 VP1 gene homology in 84.8% ~ 90.3% between SD38 strain VP1 gene and 10 strains of DHAV-1 homology between 92.2%-98.0%, SD40 and 10 strains of duck hepatitis virus type 1 VP1 gene homology in 85.9% ~ 91.8%. The homology of the 6 test strains with the VP1 gene of type 2 and type 3 duck hepatitis virus was from 62.5% to 71.6%, and the homology was low. The SD38 VP1 gene is highly homologous to the VP1 gene of 10 strains of duck hepatitis virus type 1. At the same time, the VP1 gene of SD38 virus strain was segmented in this experiment. The soluble protein pCold-TF-1Q, pCold-TF-1Z and pCold-TF-1H of high antigenicity were successfully obtained, and the obtained segmented protein was purified. This experiment laid the foundation for the establishment of ELISA detection method for detecting the antibody of type I duck viral hepatitis.
【学位授予单位】：内蒙古农业大学
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：S852.65

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