Fig. 2From: Stable expression of mucin glycoproteins GP40 and GP15 of Cryptosporidium parvum in Toxoplasma gondiiVerification 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 controlBack to article page