| 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) : 19428669
|
| 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 cl15cy1
|
| Other information parent line | A reference wild type clone from the ANKA strain of P. berghei (PubMed: PMID: 17406255). |
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| The mutant parasite was generated by |
| Name PI/Researcher | M. Tufet-Bayona, C.J. Janse, R.E. Sinden, B. Franke-Fayard |
| Name Group/Department | Leiden Malaria Research Group |
| Name Institute | Leiden University Medical Center (LUMC) |
| City | Leiden |
| Country | The Netherlands |
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| Name of the mutant parasite |
| RMgm number | RMgm-210 |
| Principal name | 452cl1 |
| Alternative name | RAP2/3-GFP |
| Standardized name | |
| Is the mutant parasite cloned after genetic modification | Yes |
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| Phenotype |
| Asexual blood stage | Analysis of GFP expression in synchronized asexual blood stage parasites in vitro showed GFP expression only in maturing schizonts, as merozoites are formed. Strong GFP expression was observed in the merozoites within the mature schizont. Merozoites showed a 'two-dot' staining pattern which is characteristic for location in the two rhoptry-organelles. |
| Gametocyte/Gamete | Not different from wild type |
| Fertilization and ookinete | Not different from wild type |
| Oocyst | RAP2/3-GFP was not expressed in gametocytes, zygotes, ookinetes and developing oocysts, up to day 9 after mosquito infection and GFP expression only appeared in sporulating oocysts (from day 9 onwards). Both oocyst-derived sporozoites and salivary gland sporozoites showed clear patterns of GFP fluorescence. In oocyst-derived sporozoites GFP fluorescence appeared as 2–6, often clearly separated, dots on the apical side of the nucleus whereas inmost (>80%) of the mature salivary gland sporozoites the GFP fluorescence was seen as a longer uniformly staining pattern beginning at the very apical end of the parasite |
| Sporozoite | Both oocyst-derived sporozoites and salivary gland sporozoites showed clear patterns of GFP fluorescence. In oocyst-derived sporozoites GFP fluorescence appeared as 2–6, often clearly separated, dots on the apical side of the nucleus whereas in most (>80%) of the mature salivary gland sporozoites the GFP fluorescence was seen as a longer uniformly staining pattern beginning at the very apical end of the parasite |
| Liver stage | Not tested |
| Additional remarks phenotype | Mutant/mutation
The mutant expresses a c-terminal GFP-tagged RAP2/3 (rhoptry-associated protein 2/3).
Protein (function)
In P. falciparum a low-molecular-weight protein complex, consisting of three non-covalently linked members, RAP1, RAP2 and RAP3 localises to the rhoptries of merozoites. RAP2 and RAP3 are encoded by adjacent genes (PFE0080c; PFE0075c), organised in a head to tail fashion on chromosome 5. Both rap2 and rap3 show a high homology with the single copy, syntenic gene, rap2/3 of P. berghei. The RAP proteins are implicated in the invasion of the erythrocyte by the merozoite.
Phenotype
The phenotype analyses indicate expression of GFP-tagged RAP2/3 in rhoptries of merozoites and rhoptries of the sporozoites. No expression was observed in ookinetes which is in agreement with the proposed lack of rhoptries in this third invasive form of Plasmodium.
Additional information
PFE0080c - rhoptry-associated protein 2, RAP2
PFE0075c - rhoptry-associated protein 3, RAP3
Gen Bank accession number P. berghei rap2/3: XM 669159.1
Other mutants
RMgm-211: Unsuccessful attempts to disrupt the rap2/3 gene |
*RMgm-210
