禽4型腺病毒knob蛋白的原核表达及纯化蛋白的免疫原性研究  

Prokaryotic Expression of Fowl Adenovirus Type 4 Knob Protein and Immunogenicity Study of Purifi ed Protein

作　　者：黄冬 卜德新 王睿智 张伯顺 于春梅 丛雁方 HUANG Dong;BU Dexin;WANG Ruizhi;ZHANG Boshun;YU Chunmei;CONG Yanfang(Qingdao Vland Biotech INC.,Qingdao 266102,China)

机构地区：[1]青岛蔚蓝生物股份有限公司,青岛266102

出　　处：《中国动物传染病学报》2023年第5期87-92,共6页Chinese Journal of Animal Infectious Diseases

摘　　要：为研究Knob蛋白的免疫效果,将合成的mknob基因克隆至pET28a(+)表达载体,转化至大肠杆菌BL21感受态细胞,低温诱导表达,knob蛋白纯化后去内毒素,按不同剂量免疫SPF鸡。结果表明:重组质粒pET28a-mknob可以高效表达目的蛋白;Western blot结果表明,表达的蛋白可与禽腺病毒特异性血清发生特异性结合;免疫保护试验分析发现,10μg蛋白免疫SPF鸡即可产生100%保护。本研究为禽4型腺病毒亚单位疫苗的研发奠定了基础。To investigate the immunological effect of Knob protein,the synthesized mknob gene was cloned into pET28a(+)expression vector,transformed into Ecoli.BL21 receptor cells,expressed at low temperature induction,and the knob protein was purified and de-endotoxified and vaccinated to SPF chickens at different doses.The results showed that the recombinant plasmid pET28a-mknob could express the target protein efficiently.Western-blot results showed that the expressed protein could be specifically bound to fowl adenovirus-specific serum.Immunoprotection assay showed that 5μg of protein could produce good protection effect when immunized with SPF chickens.This study laid a foundation for the development of Fowl adenovirus type 4 subunit vaccine.

关 键 词：禽4型腺病毒 knob蛋白 原核表达 纯化 免疫原性 

分 类 号：S852.65[农业科学—基础兽医学]