FIG 1.

Construction of transgenic Eimeria lines expressing EmPro and/or TgPro. (A) Schematic and design of recombinant vectors. Exogenous profilins and reporter EYFP were coexpressed in a single expression cassette linked by P2A. (B) Stably transfected Et-EmPro expressing the reporter EYFP in its sporozoite, schizont, and sporulated oocyst stages. Bar, 5 μm. (C) Validation of the expression of exogenous profilins by Western blotting. Parasites from sporozoite stages were immunoblotted with mouse anti-Flag tag or mouse anti-His tag monoclonal antibody. The molecular weight of EmPro with Flag tag was 20.5 kDa, that of TgPro with Flag tag was 18.9 kDa, and that of EmPro with His tag and P2A peptide was 22.6 kDa. (D) Distribution of exogenous profilins in transgenic sporozoites analyzed by IFA with mouse anti-Flag tag and mouse anti-His tag monoclonal antibody. Bar, 5 μm. (E and F) Comparison of oocyst shedding patterns (E) and reproduction (F) of the transgenic Eimeria lines and the wild type. Each value represents the mean for three birds.