Mutant/mutation
The mutant expresses a C-terminal GFP-tagged version of PBANKA_124660 under the control of the constitutive eef1a promoter (and the 3'-UTR of the calmodulin gene). The gfp-tagged gene is introduced on a circular plasmid.

Protein (function)
The protein was selected based on the presence of a PEXEL motif.

Phenotype
Expression of the GFP-tagged protein in blood stages and is exported into the red blood cell (RBC). Localization in the RBC cytoplasm (punctuate localization)

Additional information
PbCP1 harbours an atypical PEXEL motif (RxLxY) and is further characterised by two predicted transmembrane domains (2TMD) in the C-terminal end of the protein.  Evidence is presented that the 2TMD region is required to recruit PbCP1 to discrete membranous structures in the red blood cell cytosol that have a convoluted, vesico-tubular morphology by electron microscopy.

To confirm the localisation of PbCP1, a second transgenic parasite line was generated in which the GFP reporter was replaced with a triplehaemagglutinin/streptavidin tag (3xHA/Strep). IFA of acetone/methanol fixed iRBCs displayed the same export phenotype for PbCP1-3xHA/Strep as observed for the PbCP1-GFP expressing parasite line and immunoblot analysis using anti-HA antibodies revealed a doublet protein band at the predicted MW of ~30 kDa.

Other mutants