关键词: 日本血吸虫, 脂肪酸结合蛋白, 基因表达, 重组家蚕核型多角体病毒, 疫苗

Abstract: 　AIM: To express the fatty acid binding protein ( Sj14FABP) gene of Schistosoma japonicun in the silkworm cells and larvae. METHODS: A 600 bp DNA fragment containing Sj14FABP gene was cloned into baculovirus transfer vector of pBacPAK His1 to construct recombinant transfer vector Sj14-pBac PAK His1 . Coinfection was accomplished with this vector and Bombyx mori nuclear polyhedrosis virus (BmNPV) DNA in BmN cells. The recombinant virus of Bm-Sj14 was screened using dot-blotting. The BmN cells and silkworm larvae were infected with Bm-Sj14 to express Sj14FABF gene. Western blotting and ELISA were used to identify the antigenicity of the recombinant protein. RESULTS: Sj14FABP gene was successfully expressed in the BmN cells and silkworm larvae infected with Bm-Sj14. The product was a 18 kDa fusion protein. The yield in BmN cells was about 100 μg/1×10 6 cells and 33 μg/ml cell supernatant. In silkworm larvae, the product yield was 4 mg/ml haemolymph as well as 4.6 mg/g silkworm tissue. The recombinant protein could be recognized by Western blotting and ELISA using the sera from mice immunized with SWAP. CONCLUSION: Sj14FABP gene has been successfully expressed in BmNPV system and the product has high antigenicity.

Key words: Schistosoma japonicum, fatty acid binding protein, gene expression, recombinant BmNPV, vaccine