重组小瓜虫抑动抗原表达产物的纯化及其免疫学特性分析

Purification and immunological analysis of the recombinant immobilization antigen of Ichthyophthirius multifiliis

培养表达小瓜虫抑动抗原的重组四膜虫(Tetrahymena thermophila)G1株,诱导表达小瓜虫的抑动抗原,采用非离子型去污剂Triton X-114提取虫体膜蛋白,然后通过亲和层析法纯化抑动抗原。采用ELISA、SDS-PAGE和Western Blotting分析所提取的抑动抗原的纯度及免疫学特性。结果表明:通过亲和层析能够得到重组四膜虫所表达的G1抑动抗原蛋白,它与相应血清型小瓜虫抑动抗原的分子量和免疫原性相近,其分子量为44 kDa(还原条件)或36 kDa(非还原条件)。

The immobilization antigen of Ichthyophthirius multifiliis is a protective antigen of great value in protecting the endoparasite of I. multifiliis . The recombinant T. thermophila was cultured and induced to express the immobilization antigen of I. multifiliis. The non-ionic detergent, Triton X-114, was used to extract the membrane protein, and the immobilization antigen purified by affinity chromatography. The immunogenity and purity of the immobilization antigen were determined by ELISA, SDS-PAGE and the Western Blotting. The results showed that the molecular weight and immunogenity of the immobilization antigen expressed in T. thermophila were almost the same as those of the immobilization antigen of I. multifiliis. The molecular weight of G1 immobilization antigen expressed in T. thermophila was 44 kDa under reduced condition, or 36 kDa under non-reduced condition. Result of this study provided experimental data for recombinant T. thermophila that is to be used for preventing the fish from I. multifiliis infection.