Zarlenga D S, Rhoads M L, al-Yaman F M
U.S. Department of Agriculture, Biosystematic Parasitology Laboratory, Beltsville, MD 20705.
Mol Biochem Parasitol. 1994 Oct;67(2):215-23. doi: 10.1016/0166-6851(94)00132-4.
A cDNA expression library was constructed in lambda gt11 using poly A mRNA from the metacestode stage of Taenia crassiceps. The library was screened with rabbit antiserum to a previously defined protein fraction from Taenia hydatigena immunodiagnostic for bovine cysticercosis and with sera from cattle with experimentally induced cysticercosis. One clone (lambda TcA2) containing a 279-bp cDNA insert, reacted strongly with both antisera. A second clone (lambda TcA5.5) revealed the full-length cDNA sequence to be 361 bp. Data from Southern blots and enzymatically amplified genomic DNA segments were consistent with multiple copies or a gene family within the genome. The lambda TcA2 cDNA insert was subcloned into the plasmid pPR987 which generated a 47-kDa maltose-binding fusion protein (TcA2-MBP). Affinity-purified TcA2-MBP antigen reacted positively by ELISA with sera from cattle with experimentally induced T. saginata infections but not with sera from cattle with Fasciola hepatica or common gastrointestinal parasite infections. Rabbit polyclonal, monospecific antisera to TcA2-MBP recognized a 10-kDa protein in the cyst fluid, body wall and excretory/secretory products of the metacestode stage of T. crassiceps and immonolocalized this protein to organelles within the matrix of the cyst wall.
利用来自肥胖带绦虫成虫期的多聚腺苷酸mRNA，构建了λgt11 cDNA表达文库。用针对来自泡状带绦虫的先前定义的蛋白质组分的兔抗血清对该文库进行筛选，该蛋白质组分对牛囊尾蚴病具有免疫诊断作用，并使用来自实验性诱导囊尾蚴病的牛的血清进行筛选。一个含有279 bp cDNA插入片段的克隆（λTcA2）与两种抗血清均发生强烈反应。第二个克隆（λTcA5.5）显示全长cDNA序列为361 bp。Southern印迹和酶促扩增基因组DNA片段的数据与基因组内的多个拷贝或基因家族一致。将λTcA2 cDNA插入片段亚克隆到质粒pPR987中，该质粒产生了一个47 kDa的麦芽糖结合融合蛋白（TcA2-MBP）。亲和纯化的TcA2-MBP抗原通过ELISA与来自实验性诱导感染牛带绦虫的牛的血清呈阳性反应，但与来自感染肝片吸虫或常见胃肠道寄生虫的牛的血清无反应。兔抗TcA2-MBP多克隆单特异性抗血清在肥胖带绦虫成虫期的囊液、体壁和排泄/分泌产物中识别出一种10 kDa的蛋白质，并将该蛋白质免疫定位到囊壁基质内的细胞器中。
