Mutant/mutation
The mutant expresses ovalbumin (OVA) fused to Hep17/EXP1 (PY17X_0928700), a parasithophourous vacuole membrane (PVM) protein. The fusion gene ova::hep17 was under control of the hep17 regulatory 5’- and 3’-UTR sequences. The OVA::HEP17 expression cassette was introduced into into the fabb/f gene locus (PY17X_1126500) in the genome of the reference parent P. yoelii XNL line 1971cl1, that expresses the reporter fusion gene GFP-luciferase

Protein (function)
The gene encoding ovalbumin (OVA) was introduced into the fabb/f gene locus. FabB/F. FABB/F is an enzyme of the bacterial like type II fatty acid biosynthesis (FAS-II) pathway. FABB/F is not essential for blood stage development, mosquito stage development and development into maturing liver stages. It is, however, important (but not essential role) in the formation of infective liver stage merozoites.

Phenotype
Normal blood stage development/multiplication. Expression of OVA in asexual blood stages was shown by Western and immuno-fluorescence analyses. OVA::HEP17 expression at the parasitophorous vacuole membrane. We have previously shown that transgenic parasites expressing OVA fused to Hep17 induces strong OT-I and OT-II responses

Additional information
To generate transgenic P. yoelii mutant expressing ovalbumin (OVA) fused to Hep17/EXP1 of P. yoelii (PY17X_0928700) under the control of P. yoelii hep17 promoter, we generated DNA construct pL2244. First (step1), the promoter of hep17 (1.37 kb upstream of the start codon) and the signal peptide (SP) sequence of hep17 (bp 1-81) were PCR-amplified from wild-type P. yoelii genomic DNA and cloned into AflII/KpnI restriction sites of plasmid pL1980 that has sequences targeting the 3’ and 5’ regions of the fabb/f gene of P. yoelii (PY17X_1126500). Second (step 2), the remainder of the hep17 open reading frame (ORF) after the SP (bp 82-785), along with ∼800-bp of the 3′-UTR, was PCR-amplified and then cloned at MunI/KpnI restriction sites of the plasmid made in step 1. Third, the ORF of OVA, which was PCR-amplified from pL1884 was cloned into MunI/ BglI digested plasmid made in step 2. Finally, the hdhfr selectable marker (SM) cassette was amplified from pL0005 (MRA-774, deposited to the BEI Resources Repository) and cloned into KpnI-digested plasmid made in step 3, resulting in the final construct pL2244. In this construct the hdhfr SM is under control of the eef1a promote and the 3’UTR of Pbdhfr/ts. This construct was linearized using EcoRI/HindIII sites before transfection Transfection (exp. 2936), selection and cloning of transformed parasites was performed using standard genetic modification technologies using 1971cl1 as the parent P. yoelii XNL parasite line

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