Successful modification | The parasite was generated by the genetic modification |
The mutant contains the following genetic modification(s) |
Gene tagging,
Introduction of a transgene
|
Reference (PubMed-PMID number) |
Reference 1 (PMID number) : 28107409 |
MR4 number |
|
top of page |
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 507cl1 (RMgm-7)
|
Other information parent line | P.berghei ANKA 507cl1 (RMgm-7) is a reference ANKA mutant line which expresses GFP under control of a constitutive promoter. This reference line does not contain a drug-selectable marker (PubMed: PMID: 16242190). |
top of page |
The mutant parasite was generated by |
Name PI/Researcher | Koussis K; Loukeris TG; Siden-Kiamos I |
Name Group/Department | Institute of Molecular Biology and Biotechnology |
Name Institute | FORTH |
City | Heraklion |
Country | Greece |
top of page |
Name of the mutant parasite |
RMgm number | RMgm-4116 |
Principal name | PbS2P-HA |
Alternative name | |
Standardized name | |
Is the mutant parasite cloned after genetic modification | Yes |
top of page |
Phenotype |
Asexual blood stage | Localisation of PbS2PHA in proximity to the nucleus in schizonts/ merozoites and ookinetes. Double-labelling experiments with the Golgi marker ERD2 in schizonts revealed close association of S2P with ERD2, although there was not complete overlap of the signals. |
Gametocyte/Gamete | Not tested |
Fertilization and ookinete | Localisation of PbS2PHA in proximity to the nucleus in schizonts/ merozoites and ookinetes. Double-labelling experiments with the Golgi marker ERD2 in schizonts revealed close association of S2P with ERD2, although there was not complete overlap of the signals. |
Oocyst | Localisation of PbS2PHA in proximity to the nucleus in schizonts/ merozoites and ookinetes. Double-labelling experiments with the Golgi marker ERD2 in schizonts revealed close association of S2P with ERD2, although there was not complete overlap of the signals. A signal was detected in mature oocysts (d12-d14) after mosquito infection; and in mature salivary gland sporozoites. In the latter, the protease was localised close to the nucleus. |
Sporozoite | Localisation of PbS2PHA in proximity to the nucleus in schizonts/ merozoites and ookinetes. Double-labelling experiments with the Golgi marker ERD2 in schizonts revealed close association of S2P with ERD2, although there was not complete overlap of the signals. A signal was detected in mature oocysts (d12-d14) after mosquito infection; and in mature salivary gland sporozoites. In the latter, the protease was localised close to the nucleus. |
Liver stage | Not tested |
Additional remarks phenotype | Mutant/mutation
The mutant expresses a C-terminal (triple) HA-tagged version of S2P and expresses GFP under control of the eefia constitutive promoter.
Protein (function)
Plasmodium berghei encodes a single member of the M50 family of proteases (PBANKA_ 1404100), henceforth termed PbS2P.The protein is predicted to have 7 transmembrane domains and a molecular weight of 39 kDa. In silico searches in PlasmoDB (http://plasmoDB.org) and EupathDB (http://eupathdb.org) identified genes encoding M50 metalloproteases in all Plasmodium species and in most organisms of the Apicomplexa phylum, with the notable exceptions of Babesia and Cryptosporidium species. All proteins contain the characteristic HExxH motif and only one (Eimeriatenella) does not contain the NPDG motif. PbS2P has an overall amino acid identity of more than 92% with other rodent malarial parasite S2Ps; a value that drops to 70% identity when compared to human parasite orthologues and to less than 30% in S2Ps of related Apicomplexa, such as Toxoplasma and Neospora, and other eukaryotic organisms or bacteria.
Phenotype
Localisation of PbS2PHA in proximity to the nucleus in schizonts/ merozoites and ookinetes. Double-labelling experiments with the Golgi marker ERD2 in schizonts revealed close association of S2P with ERD2, although there was not complete overlap of the signals. A signal was detected in mature oocysts (d12-d14) after mosquito infection; and in mature salivary gland sporozoites. In the latter, the protease was localised close to the nucleus.
Analyses of a mutant lacking S2P (RMgm-1133) showed the following:
Reduced growth of asexual blood stages. Normal numbers of salivary gland sporozoites. Reduced infectivity of sporozoites to mice. Evidence is presented for normal liver infection but affected late liver development. Mutant liver stages in culture showed a small, albeit significant, size reduction at 48h but the number of EEFs was very similar to wild type cultures. Mutant infected hepatoma cells produced two-fold fewer merosomes as compared to wild type.
Additional information
Other mutants |