Figure 3
Strategy to generate a chimeric P. berghei parasite line expressing a PvCSP-P. vivax-like protein as additional CSP. An additional copy line in which the PvCSP-P. vivax-like CDS (PVU09738) gene was introduced into the genome as an additional copy of the gene in the neutral 230p locus. The construct that contains the ‘PvCSP-P. vivax-like gene expression cassette’ was integrated into the 230p locus on chromosome 3 of the P. berghei ANKA GIMO mother line by GIMO transfection using negative selection (5-FC), resulting in the expression of the PvCSP-P. vivax-like gene under the control of the Pbuis4 gene promoter and transcriptional terminator sequences. This construct also expresses a GFP and firefly luciferase (LUC-IAV) fusion protein under control of the constitutive Pbeef1a promoter and is selectable marker (SM)-free. The construct is integrated into the neutral p230p locus by double crossover integration. Black arrows: location of PCR primers used for the diagnostic PCR analysis.
