Fig. 1.

Construction of a recombinant E. tenella line expressing EmAMA1. a EmAMA1 with a flag tag was co-expressed with TgDHFR-EYFP and linked by P2A in the single expression cassette. b Stably transfected parasite expressing the reporter EYFP in its sporulated stage as observed by fluorescence microscopy. c The EmAMA1 gene was inserted into the genome of the transgenic parasites identified by PCR using AMA1-F1/AMA1-R2 primers, and the predicated PCR product was 1632 bp. d EmAMA1 expression in recombinant Eimeria was identified by Western blotting. The predicated size of EmAMA1 and the flag tag was 60.5 kDa. The mouse anti-EtGAPDH polyclonal antibody served as a loading control. e The exogenous EmAMA1 was primarily expressed on the cell surface and the apex of the transgenic sporozoites. The IFA experiment was conducted using the mouse anti-flag tag monoclonal antibody and the Cy3-conjugated goat anti-mouse IgG as the primary and secondary antibodies, respectively. Scale-bars: b, e, 5 μm