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Updated: Jun 9, 2026
Isolation and Characterization of Extracellular Vesicles from Adult Schistosoma japonicum
Published on: May 22, 2018
[Cloning, expression and identification of endothelial differentiation-related factor-1 gene of Schistosoma
Wang-Ping Deng1, Zheng Feng, Bin Xu
1School of Biotechnology, East China University of Science and Technology, Shanghai 200237, China.
Objective:
To clone and express endothelial differentiation-related factor (SjEDF)-1 gene of Schistosoma japonicum, analyze its immunogenicity and the stage-specific expression at different developmental stages of S. japonicum.
Methods:
Total RNA were extracted from eggs, cercariae, schistosomula and adult worms. The housekeeping gene SjActin was selected as the internal reference. According to the open reading frame for SjEDF-1 gene (GenBank accession number: AY336498), a pair of primers were designed to amplify the SjEDF-1 gene which was subcloned into pET-28a vector. The recombinant plasmid SjEDF-1/pET-28a was transformed into E. coli BL21 and induced with IPTG for expression. The recombinant protein was purified with Ni-NTA resin. The immune rabbit sera was prepared by immunizing New Zealand white rabbits with purified recombinant SjEDF-1 protein. Western blotting was used to analyze the immunogenicity and the expression level of SjEDF-1 at the different developmental stages.
Results:
The SjEDF-1 gene was detected with a band of 405 bp in eggs, schistosomula, female and male worms. The recombinant protein (rSjEDF-I) was expressed as inclusion bodies (M, 20 000). Western blotting analysis showed that the purified rSjEDF-1 protein was recognized by pooled sera of infected rabbits. The target protein was detected only in schistosomulum and adult worms.
Conclusion:
The recombinant protein (rSjEDF-I) shows certain immunogenicity, and is detected only in schistosomula and adult worms.
