Fig. 2

Verification of the correct integration and expression of the GP40 and GP15ΔGPI of Cryptosporidium parvum in Toxoplasma gondii. a Diagnostic PCR confirming the homologous integration and gene disruption in a representative clone compared to the parental line RHΔku80. PCR1 and PCR2 provide evidence of homologous integration based on products amplified between the CpGP40-Twinstrep-6 × HA-DHFR* (or CpGP15-6 × HA-DHFR*) gene and regions in the UPRT locus that lie outside of the targeting amplicon. PCR3 amplifies a 1.0-kb fragment in wild-type (WT) parasites that is lost because of the insertion of CpGP40-Twinstrep-6 × HA-DHFR* (or CpGP15Δgpi-6 × HA-DHFR*). b Expression of CpGP40-Twinstrep-6 × HA in Tguprt-Cpgp40 mutant determined by Western blotting. CpGP40 was detected with mouse anti-HA, rabbit anti-Strep, and rabbit anti-GP60, with the WT control