摘要
为进行传染性法氏囊病病毒(IBDV)多表位抗原的原核表达和免疫特性分析,应用重叠延伸PCR技术扩增合成由病毒VP2和VP3蛋白多个表位序列构成的重组抗原基因,并将抗原基因与原核表达载体pBVIL1重组,表达多表位融合抗原,通过与标准抗血清反应和动物免疫分别检测表达抗原的免疫反应性和免疫原性。结果显示,经42℃诱导后,含重组载体的大肠杆菌表达了31 kD的抗原蛋白,Western blot鉴定出现阳性条带,免疫小鼠后,抗血清的滴度为1∶6 400。说明原核表达载体构建成功,大量表达的多表位融合抗原具有IBDV抗原反应性。
To express multi-epitope antigen of infectious bursal disease virus（IBDV）and analyze its immune characteristics,the multiepitope recombinant gene resulting from VP2 and VP3 sequences was designed and amplified by PCR synthetic technology,and was cloned into pBVIL1 plasmid.The multi-epitope antigen was expressed by temperature induction at 42℃ with fusion and its immune reactivity was detected by IBDV standard antiserum,and the immunogenicity was evaluated using the titre of antiserum from mice.The results showed that multiepitope fusion antigen was expressed with 31 kD molecular weight,its immune reactivity was confirmed using standard antisera through western blot assay and the antiserum titer was 1∶6 400 with ELISA method after mice immunization.It was concluded that the prokaryotic expression vector was constructed successfully and the multi-epitope antigen possessed the IBDV antigenic properties,which could be provided for pathogen diagnosis and genetic engineering vaccine research.
出处
《生物技术通报》
CAS
CSCD
北大核心
2013年第9期165-169,共5页
Biotechnology Bulletin
关键词
传染性法式囊病毒
多表位抗原
原核表达
免疫反应性
免疫原性
Infectious bursal disease virus（IBDV） Multiple epitopes antigen Prokaryotic expression Immunogenicity Immunoreactivity
