Successful modification | The parasite was generated by the genetic modification |
The mutant contains the following genetic modification(s) |
Gene mutation,
Gene mutation,
Introduction of a transgene
|
Reference (PubMed-PMID number) |
Reference 1 (PMID number) : 29195810 |
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 |
RMgm-4110
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Other information parent line | In the mutant the endogenous P. berghei csp gene has been replaced by the P. falciparum csp gene. This has been performed by the GIMO method of transfection. The P. falciparum csp gene is under control of the 5'- and 3'-UTR regions of the P. berghei csp gene. The mutant does not contain a drug-selectable marker. The mutant also expresses the fusion protein GFP-Luciferase under control of the constitutive eefia promoter. |
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The mutant parasite was generated by |
Name PI/Researcher | Salman AM, Khan SM, Janse CJ |
Name Group/Department | Leiden Malaria Research Group |
Name Institute | Leiden University Medical Center |
City | Leiden |
Country | The Netherlands |
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Name of the mutant parasite |
RMgm number | RMgm-4987 |
Principal name | 2644cl8 cl3-6 |
Alternative name | PbANKA-PfCSP(r)PbCSP-PFTRAP(r)PbTRAP |
Standardized name | |
Is the mutant parasite cloned after genetic modification | Yes |
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Phenotype |
Asexual blood stage | Not different from wild type |
Gametocyte/Gamete | Not different from wild type |
Fertilization and ookinete | Not different from wild type |
Oocyst | Not different from wild type |
Sporozoite | Not different from wild type |
Liver stage | Not different from wild type |
Additional remarks phenotype | Mutant/mutation
In the mutant
- The endogenous P. berghei csp gene has been replaced by the P. falciparum csp gene.
- The endogenous P. berghei trap gene has been replaced by the P. falciparum trap gene
This has been performed by the GIMO method of transfection.
The mutant does not contain a drug-selectable marker. The mutant also expresses the fusion protein GFP-Luciferase under control of the constitutive eefia promoter.
Protein (function)
CSP: The CS protein is the major protein on the surface of sporozoites and is critical for development of sporozoites within the oocysts and is involved in motility and invasion of both the salivary gland of the mosquito and the liver cells. The protein is also found on the oocyst plasma membrane and on the inner surface of the oocyst capsule. Specific motifs in CS are involved in sporozoite binding to mosquito salivary glands and in sporozoite attachment to heparan sulfate proteoglycans in the liver of the mammalian host. During substrate-dependent locomotion of sporozoites, CS is secreted at the sporozoite anterior pole, translocated along the sporozoite axis and released on the substrate at the sporozoite posterior pole. Following sporozoite invasion of hepatocytes, the CS is released in the host cell cytoplasm.
TRAP: TRAP is a type 1 transmembrane protein, containing two adhesive domains in its extracellular portion, an A-domain of von Willebrand factor and a thrombospondin type I repeat (TSR, TSP). The cytoplasmic part (tail) of the protein is postulated to interact with actin–myosin motor proteins giving the force needed for motility.
TRAP is located in the micronemes of sporozoites. The protein plays a role in the gliding motility of sporozoites and invasion of host cells as has been shown by analysis of mutant parasites lacking expression of TRAP
Phenotype
Normal development throughout the complete life cycle showing complementation of P. berghei CSP by P. falciparum CSP and P. berghei TRAP by P. falciparum TRAP
Additional information
In the mutant the endogenous P. berghei trap gene has been replaced by the P. falciparum trap gene. This has been performed by the GIMO method of transfection. In the 2257cl2 parent line (expressing P. falciparum CSP) the trap open reading frame (orf) was first replaced with the hdhfr-yfcu selectable marker (SM) using plasmid pL2111 (RMgm-4112), resulting in mutant line 2560cl4. Subsequently, the SM was replaced by the P. falciparum trap orf by transfection and negative selection. The P. falciparum trap gene is under control of the 5'- and 3'-UTR regions of the P. berghei trap gene.
Other mutants |