Fig. 6
Creation of the PbclagX/loxP inducible knockout line. a Schematic of pPbclagX/flox designed to modify the PbdiCre clagX locus such that the ~ 400 aa region containing RhopH2/RhopH3 binding regions (RH2B, RH3B), the hypervariable region (HVR) and transmembrane domain (TMD) are floxed. pABR099 containing a PbclagX gRNA was utilised to cleave the PbclagX locus (lightning bolt). A schematic of the expected PbclagX/flox locus after integration of the targeting construct, and the expected truncated locus following the addition of rapamycin and loxP recombination, as is a protein schematic to highlight which domains have been lost after truncation. PbclagX/flox(192) is a clonal line of PbclagX/flox generated by serial dilution. Indicated are the primers used to validate the presence of b–d WT; e integration of loxP; f presence of cMyc; g and 3′ integration, and the expected product sizes are indicated. PbdiCre gDNA was used as a positive control for detecting WT and as a negative control for integration