鸡传染性法氏囊病病毒VP5的原核表达、多抗制备及初步应用  

Prokaryotic Expression of VP5 Protein of Infectious Bursal Disease Virus and Preparation and Preliminary Application of Its Polyclonal Antibodies

作　　者：韩金泽 牛鑫鑫 王国栋 葛成菲 张玉龙 黄萌萌 许萌萌 于航博 刘润杭 韩京哲 吴子文 于晓雪 高玉龙[2,3] 李留安 祁小乐[2,3] Han Jinze;Niu Xinxin;Wang Guodong;Ge Chengfei;Zhang Yulong;Huang Mengmeng;Xu Mengmeng;Yu Hangbo;Liu Runhang;Han Jingzhe;Wu Ziwen;Yu Xiaoxue;Gao Yulong;Li Liu'an;Qi Xiaole(College of Animal Science and Veterinary Medicine,Tianjin Agricultural University,Tianjin Key Laboratory of Agricultural Animal Breeding and Healthy Husbandry,Tianjin 300392,China;Avian Immunosuppressive Diseases Division,National Key Laboratory for Animal Disease Control and Prevention,Harbin Veterinary Research Institute,Chinese Academy of Agricultural Sciences,Harbin 150069,Heilongjiang,China;World Organization for Animal Health(WOAH)Reference Laboratory for Infectious Bursal Disease,Harbin Veterinary Research Institute,Chinese Academy of Agricultural Sciences,Harbin 150069,Heilongjiang,China)

机构地区：[1]天津农学院动物科学与动物医学学院,天津市农业动物繁育与健康养殖重点实验室,天津300392 [2]中国农业科学院哈尔滨兽医研究所,动物疫病防控全国重点实验室禽免疫抑制病创新团队,黑龙江哈尔滨150069 [3]中国农业科学院哈尔滨兽医研究所,世界动物卫生组织传染性法氏囊病参考实验室,黑龙江哈尔滨150069

出　　处：《中国动物检疫》2024年第2期92-98,共7页China Animal Health Inspection

基　　金：“十四五”黑龙江省重点研发计划项目(GA21B004);黑龙江省自然科学基金项目(ZD2020C006);国家自然科学基金项目(32072852)。

摘　　要：传染性法氏囊病(infectious bursal disease virus,IBD)是由传染性法氏囊病病毒(infectious bursal disease virus,IBDV)引起的一种主要危害雏鸡的急性传染病。在IBDV编码的5个蛋白中,VP5是IBDV唯一可以缺失基因的蛋白。本研究克隆了IBDV优势流行毒株的VP5蛋白编码基因,利用原核表达系统表达了VP5蛋白,通过免疫BALB/c小鼠制备VP5多抗,并对多抗进行了鉴定和检测不同IBDV毒株的应用。结果显示,IBDV优势流行毒株的VP5蛋白实现了可溶性表达,分子质量约为17 kDa;制备的VP5多抗特异性良好,ELISA效价可达1:64000;VP5多抗可通过IFA和Western Blot检测识别IBDV rHLJ0504-HT株,不识别VP5基因缺失株。结果表明:试验成功表达了IBDV优势流行毒株的VP5蛋白,并制备了VP5多抗;VP5多抗可对IBDV及其VP5编码基因缺失毒株进行鉴别检测。本研究对于IBDV检测方法的建立以及VP5编码基因功能的进一步研究提供了物质基础。Infectious bursal disease(IBD)is an acute infectious disease caused by infectious bursal disease virus(IBDV),mainly posing a threat to chicks.For 5 proteins encoded by IBDV genome,VP5 is the only one of which the gene may be deleted.In the study,the gene encoding VP5 protein(VP5 gene)of the dominant strain of IBDV was cloned,VP5 protein was expressed by prokaryotic expression system,and VP5 polyclonal antibodies were prepared through vaccinating BALB/c mice,then its application in various IBDV strains was identified and tested.The results showed that VP5 protein of IBDV dominant strain was expressed with solubility,and its molecular weight was about 17 kDa;the prepared VP5 polyclonal antibodies were with good specificity,and the ELISA titer could reach 1:64000;VP5 polyclonal antibodies could identify IBDV rHLJ0504-HT strain through IFA and Western Blot,but fail for VP5 gene-deleted strain.In conclusion,VP5 protein of the dominant strain of IBDV was successfully expressed,and VP5 polyclonal antibodies were prepared,which could be used to identify and test IBDV wild and VP5 gene-deleted strain.A material basis was provided by the study for the development of IBDV detection methods and further research on the function of VP5 gene.

关 键 词：传染性法氏囊病病毒 VP5 表达 多抗 应用 

分 类 号：S855.3[农业科学—临床兽医学]