鸡传染性法氏囊病病毒vp2基因在杆状病毒表达系统中的表达及免疫原性研究  

Immunogenicity of recombinant VP2 protein of infectious bursal disease virus expressed in recombinant baculoviruses

作　　者：高玉龙[1] 高宏雷[1] 王晓艳[1] 李俊山[2] 邓小芸[1] 刘伟[2] 王笑梅[1] 

机构地区：[1]中国农业科学院哈尔滨兽医研究所兽医生物技术国家重点实验室/禽传染病研究室,黑龙江哈尔滨150001 [2]东北农业大学动物医学院,黑龙江哈尔滨150030

出　　处：《中国预防兽医学报》2007年第6期427-431,共5页Chinese Journal of Preventive Veterinary Medicine

基　　金：国家973项目(2005CB523202);黑龙江省国际科技合作项目(WH05B02)

摘　　要：将鸡传染性法氏囊病病毒超强毒Gx株vp2基因克隆到载体pFastBac HTA中,构建重组转座载体pFVP2,然后将其转化DH10Bac感受态大肠杆菌,将vp2基因整合到Bacmid穿梭载体中,获得重组穿梭载体BacmidVP2;通过脂质体转染将其转染Sf9昆虫细胞,获得重组杆状病毒rBacVP2。用Western blot和间接免疫荧光试验分析表明IBDV VP2蛋白在Sf9昆虫细胞获得正确表达,所表达的重组VP2蛋白分子量约50 Ku。以rBacVP2感染Sf9细胞裂解物免疫3周龄SPF鸡,在免疫后7 d可检测到ELISA抗体;免疫后14 d可检测到琼脂免疫扩散抗体。攻毒试验表明,初次免疫后14 d对IBDV超强毒株的攻击保护率为75%,2次免疫后14 d对其的攻击保护率为100%。Recombinant transfer vector pFVP2 was constructed by cloning vvlBDV vp2 gene of the Gx isolate into pFastBac HTA. The transfer vector was transformed into E.coli DHIOBac cells that contain a baculovirus shuttle vector （bacmid） with a miniattTn7 target site and a helper plasmid. The recombinant BacmidVP2 was generated by transposing themini-Tn7 element located in pFVP2 to the mini-attTn7 attachment site on the Bacmid and then transfected into the St9 insect cells by lipofectin to produce recombinant baculovirus （rBacVP2）. Expression of IBDV VP2 in St9 cells infected with the recombinant virus was detected by Western blot and indirect immunofluorescence assay （IFA）. The lysates of rBacVP2 infected cells were used to immune 3-week-old SPF chickens. Antibodies to IBDV were detected by ELISA after 7 d post-immunization and by AGP after 14 d post-immunization. The protection rate against vvlBDV Gx infection was 75 % at 14 d post-immunization and 100 % at 14 d after the boosting immunization.

关 键 词：鸡传染性法氏囊病病毒 VP2基因 杆状病毒 免疫原性 

分 类 号：S852.659.4[农业科学—基础兽医学]