Prokaryotic expression and polyclonal antibody preparation of macrophage
migration inhibitory factor of Trichomonas vaginalis
LU Ye1, ZHU Yu-lian1， ZHU Li1, LIU Qing1， ZHU Yu-rong1, LING Wei1, TANG Chang-xia1
(1. School of Medicine, Jiangsu University, Zhenjiang Jiangsu 212013; 2. National Institute of Parasitic Diseases, Chinese Center for Disease Control and Prevention, Shanghai 200025， China）
Abstract：Objective: To prokaryotically express the macrophage migration inhibitory factor of Trichomonas vaginalis(TvMIF) and produce its polyclonal antibody in rabbits. Methods: The TvMIF gene was amplified by PCR, and cloned into pTG19T vector.Then positive gene fragment was inserted into pET30a(+) vector after doubledigesting and sequencing.The recombinant plasmid was transformed into E.coli strain BL21(DE3), and the recombinant protein TvMIF was inducibly expressed with IPTG and purified by NiNAT affinity chromatography and then immunized rabbits. The polyclonal antibody was identified by Western blotting. Results: The TvMIF gene was amplified from Trichomonas vaginalis cDNA by PCR. Squencing analysis showed that its sequence is the same as that in GenBank. The recombinant plasmid of pET30aTvMIF was constructed, and the molecular weight of the recombinant TvMIF protein was about 15.5 kDa,which was in the form of soluble protein. The polyclonal antibody of TvMIF recombinant protein was obtained, and the specific band of TvMIF was detected by Western blotting. Conclusion: This study successfully purified the MIF protein of Trichomonas vaginalis and prepared its polyclonal antibody, which will facilitate the research on the pathogenic mechanism of Trichomonas vaginalis.
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2019, Vol. 29 
