Cloning and prokaryotic expression of the ADF protein of Eimeria maxima

Objective This study sought to clone and express the Eimeria maxima ADF gene in E.coli DE3.Methods The ADF gene was amplified by RT-PCR and was then subcloned into the prokaryotic expression vector pET-28a.E.coli Rosetta(DE3) containing the recombinant plasmid pET-28a-ADF was induced by IPTG.The fusion protein was detected and identified by SDS-PAGE and Western blot.Results The recombinant expression plasmid was successfully constructed.After induction in an E.coli system,results of SDS-PAGE showed that a fusion protein of about 17 ku was successfully expressed.Results of Western blot showed that the fusion protein was readily recognized by polyclonal antibodies against E.maxima.Conclusion The recombinant expression plasmid pET-28a-ADF expressed the corresponding protein in E.coli DE3.These results should help with research on the immunogenicity of ADF.