摘要
为了解猪流行性腹泻病毒在河南流行毒株全基因组遗传进化水平,应用RT-PCR方法从疑似猪流行性腹泻病毒感染的病料中扩增出全基因组序列,并将PCR产物纯化后连接pMD20-T载体,转化JM109感受态细胞,挑选单克隆进行测序,获得的序列经过DNA STAR中Edit Sequence软件对全基因组序列进行拼接。结果表明,猪流行性腹泻病毒CH-He NXX-2017株全长28 038 nt(除去poly A);进化树分析显示,该毒株与IA1,USA Colorado2013,MN和PC22A等毒株亲缘关系较为接近,与国内外早期分离株如CV777,LZC,DR13等毒株亲缘关系较远。揭示猪流行性腹泻病毒一直处于不断的进化中,需要一直密切监控病毒进化趋势,为临床诊断以及疫苗研发等提供理论参考。
To understand the evolution of porcine epidemic diarrhea virus circulating in Henan province from full genome sequence level,RT-PCR was employed to amplify the full genome sequence from porcine epidemic diarrhea suspicious samples.The PCR products were purified and ligated with pMD20-T simple vector,transformed into competent cell JM109.The sequences were then abbembled using Edit Seq of DNA STAR software.The results showed that the full length of CH-He NXX-2017 was 28 038 nt in length.Phylogenetic analysis revealed that CH-He NXX-2017 clustered most closely to IA1,USA Colorado 2013,MN and PC22 A strain,far away to early isolated strain,such as CV777,LZC,DR13.The results indicated that PEDV has been evolving under immune press,attention should be paid to monitor the trend of PEDV evolving,thus providing information for clinical diagnosis and cvaccine development.
作者
焦文强
许峰
徐引弟
王治方
张青娴
王克领
朱文豪
李海利
张立宪
郎利敏
JIAO Wenqiang;XU Feng;XU Yindi;WANG Zhifang;ZHANG Qingxian;WANG Keling;ZHU Wenhao;LI Haili;ZHANG Lixian;LANG Limin(Institute of Animal Husbandary and Veterinary,Henan Academy of Agricultural Sciences,Zhengzhou 450002,China;Henan Key Laboratory of Farm Animal Breeding and Nutritutional Regulation,Zhengzhou 450002,China)
出处
《山西农业科学》
2019年第1期113-116,120,共5页
Journal of Shanxi Agricultural Sciences
基金
河南省农业科学院自主创新项目(2018ZC49)
