关键词: 日本血吸虫, IgE, 基因克隆, 重组抗原, 融合蛋白

Abstract: 　Objective To obtain the gene encoding specific IgE-related antigen of Schistosoma japonicum from the cDNA library of adult Schistosoma japonicum . Methods The pooled sera from 15 individuals with high levels of specific IgE antibody against SWAP were absorbed with Protein-G and used for screening of IgE-related antigen from the adult worm cDNA library of Schistosoma japonicum . The inserted cDNA was amplified by PCR and sequenced. According to the first reading frame of the sequence, a pair of new probes, in which Eco R Ⅰ and Not Ⅰsites were incorporated respectively,were designed and used to amplify the target gene. Then,the gene was cloned into vector pGEM-T and subcloned into expression vector pGEX-6p-1. The fusion protein was expressed, analysed by SDS-PAGE and identified by Western blotting with the specific IgE antibody, respectively. Results The inserted cDNA fragment from the positive clone was about 1 200 bp,with the ORF of 507 bp which encoded 169 amino acids. The deduced molecular weight of the recombinant protein was 19.3 kDa. The homology between the target gene(Sj43B) and other known DNA sequences was less than 40%. The fusion protein expressed by the recombinant vector pGEX-6p-1/Sj43B could be recognized by schistosome specific IgE antibody. Conclusion Sj43B may encode the specific IgE-related antigen of Schistosoma japonicum . The successful construction of recombinant plasmid pGEX-6p-1/Sj43B lay the groundwork for further studies on immunological characteristics and protection immunity of the recombinant protein.

Key words: Schistosoma japonicum, IgE, gene cloning, recombinant antigen, fusion protein