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目的 :用核酸原位杂交技术观察细粒棘球绦虫 6 6kDa抗原mRNA在虫体上的细胞定位以及在虫体发育的不同时期mRNA含量的变化。方法 :以 6 6kDa抗原的cDNA克隆EgA31为模板制备RNA探针 ,与细粒棘球绦虫原头蚴、育囊和成虫虫体进行原位杂交实验。结果 :原位杂交实验显示EgA31mRNA主要位于原头蚴和成虫的皮层下某种细胞中 ,并在棘球蚴育囊的生发层中也存在这种细胞。在原头蚴向成虫的发育过程中虫体吸盘处细胞内EgA31mRNA含量显著增加。结论 :含EgA31mRNA的细胞很可能是皮层细胞 ;EgA31蛋白可能参与吸盘上肌纤维的收缩作用并与虫体的免疫逃避功能有关。
OBJECTIVE: To observe the cellular localization of 6 6 kDa antigen of Echinococcus granulosus and the changes of mRNA levels in different stages of development of the worms by nucleic acid in situ hybridization. Methods: The RNA probe was prepared by using the cDNA clone EgA31 of 6 6 kDa as a template. In situ hybridization was carried out with Echinococcus granulosus, cysts and adult worms of Echinococcus granulosus. Results: In situ hybridization showed that EgA31 mRNA was located mainly in some cells under the cortex of protozoan and adult cells, and was found in the germinal layer of echinocystic cyst. EgA31mRNA expression in the sucker of the parasitoid increased significantly during the development of the protoscoid. CONCLUSION: EgA31 mRNA-containing cells are likely to be cortical cells. EgA31 protein may be involved in the contraction of muscle fibers on the sucker and related to the immune evasion function of parasites.