日本血吸虫C1q结合蛋白基因的克隆表达及其免疫保护效果评估  

CLONING AND EXPRESSION OF C1Q BINDING PROTEIN GENE OF SCHISTOSOMA JAPONICUM AND ASSESSMENT OF ITS IMMUNE PROTECTION

作　　者：贾秉光 洪炀[1] 韩倩[1] 吕超[1] 王涛[1] 柴淑梅 曹晓丹[1] 陆珂[1] 宰金丽 马帅[1] 林矫矫[1,2] 傅志强[1] 

机构地区：[1]中国农业科学院上海兽医研究所农业部动物寄生虫学重点开放实验室,上海200241 [2]江苏省动物重要疾病与人兽共患病防控协同创新中心,扬州225009

出　　处：《中国动物传染病学报》2016年第3期66-71,共6页Chinese Journal of Animal Infectious Diseases

基　　金：国家自然科学基金(31472188);国家科技支撑计划(2015BAI09B04);上海市领军人才后备队计划(201442)

摘　　要：本研究利用PCR技术扩增到日本血吸虫C1q BP基因,构建了重组表达质粒p ET-32a(+)-Sj C1q BP,并在大肠杆菌中进行表达。将重组蛋白免疫小鼠评估其免疫保护效果,采用Western blot检测其免疫原性,ELISA检测其特异性抗体水平,应用补体溶血试验分析重组蛋白对补体溶血的抑制效果。结果显示日本血吸虫C1q BP基因含729 bp的编码序列,在大肠杆菌中BL21(DE3)中表达后,用His-Bind亲和层析纯化可获得50 k Da的重组蛋白r Sj C1q BP。应用重组蛋白r Sj C1q BP免疫BALB/c小鼠可产生较高水平的特异性Ig G抗体,诱导36.08%的减虫率和43.45%的肝脏减卵率;补体溶血试验结果表明该重组蛋白能够抑制补体溶血。本研究结果为进一步研究日本血吸虫Sj C1q BP的生物学功能奠定了基础。The ClqBP gene ofS.japonicum （SjClqBP） was cloned, expressed and characterized in the present study. The open reading frame （ORF） of SjClqBP consisting of 729 bp encoding 242 amino acids was successfully expressed in Escherichia coli BL21 （DE3）. The Ni-NTA purification system was applied to purify the recombinant protein and a 50 kDa purified protein was obtained. Immunization with the purified rSjC lqBP emulsified ISA206 adjuvant induced a higher level of specific IgG and significant reductions in worms burden （36.08%） and liver eggs number （43.45%）. Hemolytic testing indicated that rSjClqBP inhibited the process of complement hemolysis. These findings suggested that SjC1 qBP might be a potential vaccine candidate against S.japonicum and laid a path for further research on its biological function.

关 键 词：日本血吸虫 C1qBP基因 补体 重组蛋白 

分 类 号：S852.735[农业科学—基础兽医学]