1型鸭甲肝病毒LY0801株VP1基因在鸭胚传代中的变异规律研究.pdfVIP

中国动物传染病学报 2014,22(5): 21-27 Chinese Journal of Animal Infectious Diseases ·研究论文· 1 型鸭甲肝病毒 LY0801 株 VP1 基因在鸭胚传代中 的变异规律研究 马明杰，郭翠平，焦玉祥，吕佳泓，孙 振，张瑞华，陈君豪，谢之景，姜世金 （山东农业大学动物医学院山东省动物生物工程与疾病防治重点实验室，泰安 271018 ） 摘 要：为了解1型鸭甲肝病毒（Duck hepatitis A virus type 1 ）VP1基因在鸭胚传代中的变异规律，本研究将LY0801株DHAV-1在 鸭胚体内传代至30代，分别对1~5、10、15、20 、25 、30代病毒进行VP1基因的克隆测序。各代次病毒分别以108copies/胚接种9 日龄健康鸭胚，测定各组鸭胚的平均死亡时间和病毒在鸭胚尿囊液中的增殖拷贝数。结果表明：DHAV-1在鸭胚传代过程中， 不同代次之间出现12个氨基酸的反复变异和同步变异，分别为R43M （K ）、T48A、T101S、L169F 、T180I、S181L、R183Q 、 E184A （K ）、G187D、D193N 、M213R 、H219Y ；在传代过程中，病毒致死鸭胚的时间逐渐延迟，但病毒在鸭胚内的增殖拷 贝数未呈现稳定增长的趋势。 关键词：1型鸭甲肝病毒；VP1 ；变异；致弱 中图分类号：S852.659.6 文献标志码：A 文章编号：1674-6422 （2014 ）05-0021-07 GENETIC VARIATION OF VP1 GENE OF DUCK HEPATITIS A VIRUS TYPE 1 LY0801 STRAIN DURING REPEATED PASSAGES IN DUCK EMBRYOS MA Ming-jie, GUO Cui-ping, JIAO Yu-xiang, LV Jia-hong, SUN Zhen, ZHANG Rui-hua, CHEN Jun-hao, XIE Zhi-jing, JIANG Shi-jin (Shandong Province Key Laboratory of Animal Biotechnology and Disease Control and Prevention, College of Veterinary Medicine, Shandong Agricultural University, Taian 271018, China) Abstract: To survey the genetic variation of VP1 gene of Duck hepatitis A virus type I (DHAV-I), DHAV-1 LY0801 isolate was carried 30 serial passages in duck embryos. The VP1 gene of the fi rst to fi fth, tenth, fi fteenth, twentieth, twenty-fi fth and thirtieth passages was amplifi ed and sequenced respectively. With the dose of 108 copies per embryo, each ten 9-day-old healthy duck embryos were inoculated with different passages of virus, and the mean death time of each group and the vial copies in duck embryo allantoic fl uid were detected. The results showed that there were 12 amino acid mutation sites in VP1 gene in different passages, which were R43M(K), T48A, T1