Fig. 2

Verification of stable Emi.chFc population. a Fluorescent images of the fifth generation of Emi.chFc oocysts. b Detection of chFc and YFP genes by conventional PCR. Lanes 1 and 5, PCR products from plasmid pMDEAAsschFcA (positive control); Lanes 2 and 6, Nuclear DNA extracted from transgenic E.mitis; Lanes 3 and 4, Nuclear DNA extracted from wild type E. mitis (negative control); M: DNA marker (DL 2000 plus marker). c Agarose gel electrophoresis of amplified products of PCR-based genome walking. 1–3 represents the number of PCR runs. AP2 is a random forward primer supplied in the Genome walking kit. M: DNA marker (DL2000 plus marker). d Integration sites of the expression cassette in the E.mitis genome detected using the Genome walking technique. e Western blotting detection of recombinant chFc protein. Lane1 and lane 2: SDS-PAGE immunoblot of transgenic and wild type E. mitisproteins, respectively. f Cellular localization patterns of chicken Fc in the sporozoites of E. mitis by IFA. Scale-bar: 10 μm