机构地区: 中国农业科学院哈尔滨兽医研究所
出 处: 《中国免疫学杂志》 2005年第2期127-133,共7页
摘 要: 目的 :研究猪瘟病毒 (CSFV)E2蛋白重复多抗原表位基因的融合表达及其免疫攻毒保护作用。方法 :应用PCR方法扩增猪瘟病毒E2蛋白重复多抗原表位基因 ,构建重复多表位基因的原核重组表达质粒PGEX-3E ,进行融合蛋白的表达和纯化。ELISA和Westernblot方法测定单表位融合蛋白GST-E和多表位融合蛋白GST-3E与猪抗CSFV的阳性血清和兔抗E2阳性血清的反应性 ,并进行融合蛋白的兔体免疫及免疫攻毒保护的比较研究。结果 :分子克隆和构建了原核重组表达质粒pGEX-3E ,表达和纯化了融合蛋白GST-3E ;单表位融合蛋白与重复多表位融合蛋白均能够与猪抗CSFV的阳性血清反应和兔抗E2的阳性血清产生免疫反应。在刺激兔体产生抗体方面 ,单表位融合蛋白刺激兔体产生抗体的能力较弱 ,而多表位融合蛋白则能够使兔体产生高效价的抗体。接种 10 0MID50 剂量猪瘟兔化弱毒 (HCLV)进行的兔体免疫攻毒保护试验表明 ,空白对照组和载体蛋白GST免疫组无保护作用 ,单表位融合蛋白免疫组具有一定的免疫保护作用 ,而重复多表位融合蛋白免疫组则完全能够抵抗猪瘟病毒的攻击。结论 :猪瘟病毒E2蛋白重复多表位的融合表达具有免疫保护作用 。 Objective:To investigate the fusion expression of multiepitope gene of CSFV envelope glycoprotein E2 and its protection efficiency against homologous challenge in rabbits.Methods:The multiepitope gene of CSFV envelope glycoprotein E2 was amplified by PCR technique.The PCR product was cloned into pGEX-3X vector,then transformed into E.coli BL21 cells and expressed.The single epitope fusion protein (GST-E) and multieptope fusion protein (GST-3E) were analyzed by ELISA and Western blot with pig serum against CSFV and rabbit serum against E2.The protection efficiency of the GST-E and GST-3E was determined by challenge of hog cholera lapinized virus (HCLV).Results:The GST-E and GST-3E can interacted with pig serum against CSFV and rabbit serum against E2.The capacity of inducing antibody of GST-E by immunizing rabbit is low,but GST-3E is high.The rabbits challenged with 100 MID_(50) HCLV demonstrated that GST-E immunized rabbits showed a mild increase of body temperature,GST-3E immunized rabbits no body temperature increasing on the other hand the GST immunized and control rabbits body temperature increased and then recover normally.Conclusion:The GST-3E can induce an efficient immune protection against HCLV infection,and might provide a new kind of vaccine for CSFV.