摘要
VP2蛋白是鸡传染性法氏囊病病毒(IBDV)的主要结构蛋白及保护性抗原,为了研究原核表达的IBDV VP2可溶性蛋白的免疫原性,从IBDV超强毒SD株克隆出VP2基因,将其克隆于pET-30a原核表达质粒,并转化至大肠杆菌菌株BL21中,诱导表达后,经SDS-PAGE及Western blot分析,发现在37 kDa处出现特异性蛋白条带,与预期结果一致,且大部分VP2蛋白存在于菌液上清液中。经诱导条件的优化,菌液上清液表达的VP2蛋白琼脂扩散(AGP)效价可达1∶16。用不同AGP效价的VP2蛋白乳化配苗,免疫SPF鸡,免疫后21 d,进行AGP抗体效价测定及攻毒保护试验,发现VP2蛋白以AGP效价不低于1∶8配苗,免疫鸡用超强毒SD株及经典株BC6/85株攻毒,保护率均为10/10。结果表明,成功获得了表达VP2可溶性蛋白的基因工程表达菌BL21-VP2,且表达的VP2蛋白具有良好的免疫原性,为IBDV基因工程亚单位疫苗的研制奠定了基础。
VP2 protein is a major structural protein and protective antigen of Infectious bursal disease virus(IBDV).In order to study immunogenicity of the IBDV VP2 soluble protein,the VP2 gene from the very virulent IBDV SD strain was cloned into the plasmid pET-30a and transformed into the E.coli BL21 for prokaryotic expression.The expressed VP2 protein was examined in SDS-PAGE and Western blot.The results showed that a specifi c protein band at 37 kDa was obtained,which was consistent with the expectation.Most of VP2 protein was found in supernatant in bacterial culture.After optimization of induction conditions,the agar diffusion experiment(AGP)titer of VP2 protein in bacterial supernatant reached to 1∶16.The VP2 protein with different AGP titers was emulsifi ed to prepare the vaccine for immunization of SPF chicken s.Then the titers of AGP antibodies were determined and chickens were challenged at 21 days post immunization.The chickens immunized with the VP2 vaccine produced antibodies with AGP titer of no less than 1∶8.All 10 immunized chickens were protected against challenges with the very virulent SD strain and classical BC6/85 strain.In summary,soluble BL21-VP2 protein from prokaryotic expression had good immunogenicity,which laid a solid foundation for the development of IBDV subunit vaccine.
作者
肖倩
黄秀英
丁莉
左榕琳
李成山
邢刚
XIAO Qian;HUANG Xiuying;DING Li;ZUO Ronglin;LI Chengshan;XING Gang(Chengdu Shiji Biological Products co.,ltd.,Chengdu 610100,China;Beijing Yu Kou Poultry Limited Liability Company,Beijing 101206,China)
出处
《中国动物传染病学报》
CAS
北大核心
2022年第4期156-162,共7页
Chinese Journal of Animal Infectious Diseases
作者简介
肖倩,女,硕士,主要从事禽用疫苗的研发工作;通信作者:李成山,E-mail:lichengshan521@163.com;通信作者:邢刚,E-mail:galenxing@aliyun.com。
