非洲猪瘟病毒p72蛋白的真核表达及反应原性鉴定  

Eucarytotic Expression and Reactogenicity Identification of African Swine Fever Protein p72

作　　者：梁云浩[1,2] 曹琛福[1] 叶奕优[1] 花群义[1] 张彩虹[1] 曾少灵[1] 陶虹[1] 廖立珊[1] 刘建利[1] 唐勇[2] 吕建强[1] 

机构地区：[1]深圳出入境检验检疫局动植物检验检疫技术中心,广东深圳518045 [2]暨南大学广东省分子免疫与抗体工程重点实验室,广东广州510632

出　　处：《动物医学进展》2015年第6期29-33,共5页Progress In Veterinary Medicine

基　　金：国家高技术研究发展计划项目(2012AA101301);农业公益性行业专项(200903037)

摘　　要：将p72基因克隆到载体pFast Bac/NT-TOPO上,转化大肠埃希菌DH5α。将获得的重组载体pFastBac/NT-TOPO-p72,转化大肠埃希菌DH10Bac,获得重组杆状病毒质粒bacmid-p72。通过脂质体介导,将bacmid-p72转染sf9细胞,获得重组杆状病毒。用重组杆状病毒感染正常生长的sf9细胞,72h后收集细胞,超声破碎后取上清进行SDS-PAGE和Western blot分析。结果显示,重组蛋白大小约78ku,且重组蛋白可以与非洲猪瘟标准阳性血清反应。说明表达的重组蛋白为可溶性的,并具有良好的反应原性。In order to express African swine fever virus protein p72, the p72 gene was cloned into pFastBac/ NT-TOPO vector, and then transformed into E. coil DHSa to obtain recombinant vector pFastBac/NT- TOPO-p72. Next, the pFastBac/NT-TOPO-p72 was transformed into E. coil DH10 Bac to get recombi- nant bacmid-p72. The recombianant baculovirus was generated by lipsome mediation. The sf9 cells were infected by recombinant baculovirus, the cells were collected after 72 h. The supernant was analyzed by SDS-PAGE and Western-blot after ultrasonic breaking. The result showed that the recombinant protein was about 78 ku, and could react with African swine fever standard positive serum. The results above demonstrated that, the expressed recombinant protein was soluble, and has good reactogenicity.

关 键 词：非洲猪瘟病毒 p72蛋白 脂质体介导 重组杆状病毒 

分 类 号：S852.651[农业科学—基础兽医学]