兔豆状囊尾蚴半胱氨酸蛋白酶基因的表达及免疫保护性分析

Cloning,Expression and Analysis of the Iimmunoprotection Activity of the Cysteine Protease fromTaenia pisiformis

为探讨豆状囊尾蚴半胱氨酸蛋白酶的免疫保护性能,在毕赤酵母中表达半胱氨酸蛋白酶基因,表达产物经分子筛纯化后免疫试验家兔,3免之后进行人工攻虫保护试验,且以豆状囊尾蚴烯醇酶为对照进行同步免疫和攻虫试验。结果显示,成功表达了半胱氨酸蛋白酶蛋白,且该蛋白能够识别兔豆状囊尾蚴病阳性血清;半胱氨酸蛋白酶蛋白和豆状囊尾蚴烯醇酶抑虫效果均较差。结果表明,酶蛋白在免疫保护方面的功能比较弱,不适于做疫苗的候选抗原。

To investigate the immunogenicity of cathepsin L-like cysteine protease （TpCP） ,the TpCP gene sequence was cloned from T .pisiformis metacestodes and expressed in yeast .West-ern blot analysis showed that the recombinant protein could be recognized by the positive sera of rabbit against the whole Cysticercus pisiformis .Rabbits were immunized with the expressed pro-tein that was purified by molecular sieve and feed eggs after determining antibody titer .The eno-lase of Cysticercus pisiformis was selected for further evaluating the possibility of zymoprotein as vaccine .The results showed that the immunoprotections induced by these two proteins were weak and unfit for vaccine preparation .