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Updated: Aug 29, 2026

Functional Cloning Using a Xenopus Oocyte Expression System
Published on: January 30, 2016
[Cloning, expressing and functional analysis of SjMF4, a novel Schistosoma japonicum gene]
Liang Zhang1, Guo-Feng Cheng, Zhi-Qiang Fu
1Shanghai Institute of Animal Parasitology, the Chinese Academy of Agricultural Science, Shanghai 200232, China.
Abstract:
Based on the phenomenon of the natural anti-schistosomiasis in Microtus fortis, the sera from normal Microtus fortis were employed to immunoscreen the cDNA library of adult Schistosoma japonicum (Chinese strain), two positive clones were obtained, RACE technique was further applied to amplify one of the clones, and a cDNA fragment with an ORF was identified. Sequencing revealed that it was a novel gene of Schistosoma japonicum, and it was named SjMF4 (Schistosoma japonicum Microtus fortis 4). Then the structure and functional motifs of SjMF4 were analysed. The gene was subcloned into pET-28a(+) vector; the recombinant protein showed good antigenicity in Western blotting. The gene was further subcloned into eukaryotic expression vector pcDNA3 to construct the DNA vaccine containing SjMF4. Immune experiments in mice showed significant protection that the recombinant plasmid did induce 28.64%+/-3.82% worm reduction and 21.73%+/-3.98% egg reduction than controls against the Schistosoma japonicum cercaria challenge.
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