论文摘要
目的构建中国大陆株细粒棘球蚴（Echinococcus granulosus Eg）铁蛋白（ferritin）基因的重组表达质粒,表达纯化重组铁蛋白,并对重组铁蛋白进行免疫学特性鉴定及免疫保护力研究,为包虫病分子疫苗的研制提供新的候选分子。方法（1）重组Eg.ferritin/pET-28a表达质粒的构建、表达、免疫学特性鉴定;①将目的基因亚克隆于pET-28a表达载体,转化入BL21（DE3）plysS,诱导表达重组铁蛋白（rEg.ferritin）,经含Ni2+的His-bind树脂柱纯化rEg.ferritin;②用rEg.ferritin免疫小鼠制备抗血清,通过Western-blot及ELISA等方法对rEg.ferritin的免疫学特性进行鉴定;（2） rEg.ferritin诱导小鼠免疫保护力研究:用细粒棘球蚴的原头蚴腹腔攻击感染被rEg.ferritin免疫后的小鼠,经5个月后剖杀小鼠进行免疫保护力研究。结果（1）成功构建了rEg.ferritin/pET-28a /BL21（DE3）plysS基因工程菌株,表达并纯化出rEg.ferritin。Western-blot检测:rEg.ferritin免疫小鼠的抗血清可识别rEg.ferritiney及天然抗原（原头蚴）中的Eg.ferritin,其位置大致相同约19kD。ELISA结果表明:免疫组血清中的IgG值明显高于对照组血清IgG值（P<0.01）。（2） rEg.ferritin能诱导小鼠产生98.2%的免疫保护力;结论（1）成功的构建了rEg.ferritin/pET-28a /BL21（DE3）plysS基因工程菌株并纯化制备了重组的ferritin,免疫学鉴定初步说明rEg.ferritin有较好的抗原性及免疫原性;（2） rEg.ferritin能诱导小鼠产生98.2%免疫保护力,表明rEg.ferritin具有作为包虫疫苗候选分子的潜能。
论文目录
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