表达传染性腔上囊病病毒VP2蛋白重组乳酸菌的构建及其免疫保护效果  

Construction and immunoprotective effect of recombinant lactobacilli expressing VP2 protein of infectious bursal disease virus

作　　者：齐炳理[1] 葛俊伟[1] 乔薪瑗[1] 姜艳萍[1] 李一经[1] 

机构地区：[1]东北农业大学动物医学院,黑龙江哈尔滨150030

出　　处：《中国兽医科学》2009年第11期978-983,共6页Chinese Veterinary Science

摘　　要：为探讨表达传染性腔上囊病病毒(IBDV)VP2蛋白重组乳酸菌的口服免疫效果,采用RT-PCR方法扩增了1株IBDV流行毒株的VP2基因,再分别亚克隆到干酪乳杆菌和乳酸乳球菌的表达载体pPG2和pNZ8112中,并分别电转化干酪乳杆菌L.casei393和乳酸乳球菌NZ9000的感受态细胞,构建了重组菌pPG2-VP2/L.casei393和pNZ8112-VP2/NZ9000。分别经20 g/L乳糖和1 ng/mL的乳链菌肽诱导表达后,对菌体裂解物进行了SDS-PAGE和Western-blot分析。结果显示,重组菌pPG2-VP2/L.casei393和pNZ8112-VP2/NZ9000的裂解物中分别出现大小约为53 ku和50 ku的反应带,大小均与预期结果相符,表明目的蛋白在重组菌中得到了表达。重组菌pPG2-VP2/L.casei393和pNZ8112-VP2/NZ9000分别口服免疫SPF雏鸡后,均刺激机体产生了特异性抗IBDV VP2蛋白的血清抗体;攻毒保护试验结果显示,其保护指数分别为62.5%和37.5%,表明重组菌免疫雏鸡后,对IBDV的攻击均具有不同程度的免疫保护作用。To explore the protective efficacy of recombinant lactobacillus expressing VP2 protein of infectious bursal disease virus（IBDV） through oral inoculation against IBD,Lactobacillus casei393 and Lactococcus lactis NZ9000 was transformed with segments of VP2 inserted into the pPG2 and pNZ8112 surface-expression vectors,and induced with 20g/L lactose and 1ng/mL nisin respectively,and VP2 expression was analyzed by SDS-PAGE and Western-blot.About 53ku and 50ku ladders were detected respectively from the cell lysates of pPG2-VP2/L.casei393 and pNZ8112-VP2/NZ9000 in Western-blot,indicating that the interest protein was expressed in the two different lactobacillus expression systems.SPF chicks were immunized orally with pPG2-VP2/L.casei393 or pNZ8112-VP2/NZ9000,and systemic humoral immune responses were examined.Specific anti-VP2 IgG was detected in serum.Furthermore,the protection index of pPG2-VP2/L.casei393 and pNZ8112-VP2/NZ9000 were 62.5% and 37.5% respectively,indicating that the two recombinants had immunoprotective effect in immunized chickens to different degrees.The results suggested that oral immunization with VP2-expressing Lactobacillus could induce immune response against IBDV infection.

关 键 词：传染性腔上囊病病毒 VP2蛋白 干酪乳杆菌 乳酸乳球菌 免疫 

分 类 号：S852.659.4[农业科学—基础兽医学]