| Successful modification | The parasite was generated by the genetic modification |
| The mutant contains the following genetic modification(s) |
Gene tagging
|
| Reference (PubMed-PMID number) |
Reference 1 (PMID number) : 25454088
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| MR4 number |
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| Parent parasite used to introduce the genetic modification |
| Rodent Malaria Parasite | P. berghei |
| Parent strain/line | P. berghei ANKA |
| Name parent line/clone |
P. berghei ANKA 2.34
|
| Other information parent line | P. berghei ANKA 2.34 is a cloned, gametocyte producer line of the ANKA strain (PubMed: PMID: 15137943). |
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| The mutant parasite was generated by |
| Name PI/Researcher | Sala, KA; Blagborough AM |
| Name Group/Department | Department of Life Sciences |
| Name Institute | Sir Alexander Fleming Building, Imperial College |
| City | London |
| Country | UK |
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| Name of the mutant parasite |
| RMgm number | RMgm-1150 |
| Principal name | PbPSOP12-EGFP |
| Alternative name | |
| Standardized name | |
| Is the mutant parasite cloned after genetic modification | No |
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| Phenotype |
| Asexual blood stage | Not different from wild type |
| Gametocyte/Gamete | Expression of the GFP-tagged PSOP12 exclusively in gametocytes, gametes and ookinetes. |
| Fertilization and ookinete | Expression of the GFP-tagged PSOP12 exclusively in gametocytes, gametes and ookinetes. |
| Oocyst | Not different from wild type |
| Sporozoite | Not different from wild type |
| Liver stage | Not tested |
| Additional remarks phenotype | Mutant/mutation
The mutant expresses a C-terminal GFP-tagged version of PSOP12
Protein (function)
The protein was detected in a proteome analysis of ookinetes and contains a predicted signal sequence. PSOP12 is a putative member of the well-characterized 6-Cys family of proteins. PSOP12 has been previously identified in a reverse genetics screen and medium-throughput knockout study in P. berghei, and was demonstrated as expressed in the ookinete. Expression of the protein is predicted in both the P. berghei gametocyte and ookinete. A signal peptide is located at amino acids 1–24.
Multiple gene knockouts of psop12 (RMgm-89, RMgm-254) have resulted in no detectable phenotype in blood-, sexual-, liver or pre-erythrocytic stages, suggesting that the protein could have a non-essential function within the parasite lifecycle.
Phenotype
Expression of the GFP-tagged PSOP12 exclusively in gametocytes, gametes and ookinetes
Additional information
In all stages where expression was observed much of the fluorescence signal emanated from the cell periphery, suggesting localization to the plasmalemma. This was further confirmed by IFA on fixed parasites performed with anti-EGFP mAb under non-permeablizing conditions,which demonstrated surface staining in the gametes and ookinetes. Identical surface staining was noted in transgenic PbPSOP12-EGFPparasites, and in WT P. berghei parasites stained with anti-BDES-PbPSOP12-spider serum, confirming the localization of PbPSOP12on the surface of the sexual/ookinete stages of the parasite, whilst suggesting that the presence of the EGFP-fusion protein in the transgenic line does not alter the native localization of the protein.Staining in the zygote was not observed.
In this study it was shown that immunisation of mice with PSOP12 (expressed in the baculovirus dual expression system) induces anti-malarial transmission blocking immunity.
Other mutants
Multiple gene knockouts of psop12 (RMgm-89, RMgm-254) have resulted in no detectable phenotype in blood-, sexual-, liver or pre-erythrocytic stages |
*RMgm-1150
