Back to search resultsSummaryRMgm-4158
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Successful modification | The parasite was generated by the genetic modification |
The mutant contains the following genetic modification(s) | Gene mutation, Gene tagging |
Reference (PubMed-PMID number) |
Reference 1 (PMID number) : 28481199 |
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-4155 |
Other information parent line | A drug selectable marker free P. berghei (ANKA 2.34) mutant that lakcs the cdpk4 gene. |
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The mutant parasite was generated by | |
Name PI/Researcher | Fang H, Brochet M |
Name Group/Department | Department of Microbiology and Molecular Medicine |
Name Institute | University of Geneva |
City | Geneva |
Country | Switzerland |
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Name of the mutant parasite | |
RMgm number | RMgm-4158 |
Principal name | CDPK4(G2A)-2xmyc |
Alternative name | |
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 | No exflagellation (male gamete formation). Reduced DNA replication during male gamete formation (mainly blocked at 1N). Normal mitotoic spindle formation in microgametocytes at 10 min. after activation. Strongly reduced axoneme formation in microgametocytes at 10 min. after activation. |
Fertilization and ookinete | Not different from wild type |
Oocyst | Not tested |
Sporozoite | Not tested |
Liver stage | Not tested |
Additional remarks phenotype | Mutant/mutation Protein (function) CDPK4 plays a role in male gamete formation after activation of male gametes. It plays a role in male gametocytes axoneme formation, formation of mitotic spindles and DNA synthesis. Evidence has been presented for an additional role in sporozoites: reduced invasion of hepatocytes (RMgm-1510). See PF3D7_0717500 for other CDPK4 mutants The mutant expressing the mutated CDPK4G2A form showed the following phenotype: CDPK4 has previously been shown to be myristoylated in P. falciparum. In the mutated form the myristoylated glycine 2 was replaced by an alanine residue preventing myristoylation of CDPK4. |
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Details of the target gene | |||||||||||||||||||||||||||
Gene Model of Rodent Parasite | PBANKA_0615200 | ||||||||||||||||||||||||||
Gene Model P. falciparum ortholog | PF3D7_0717500 | ||||||||||||||||||||||||||
Gene product | calcium-dependent protein kinase 4 | ||||||||||||||||||||||||||
Gene product: Alternative name | CDPK4 | ||||||||||||||||||||||||||
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Details of the genetic modification | |||||||||||||||||||||||||||
Short description of the mutation | myristoylated glycine 2 was replaced by an alanine residue preventing myristoylation of CDPK4 | ||||||||||||||||||||||||||
Inducable system used | No | ||||||||||||||||||||||||||
Short description of the conditional mutagenesis | Not available | ||||||||||||||||||||||||||
Additional remarks inducable system | |||||||||||||||||||||||||||
Type of plasmid/construct | (Linear) plasmid double cross-over | ||||||||||||||||||||||||||
PlasmoGEM (Sanger) construct/vector used | No | ||||||||||||||||||||||||||
Modified PlasmoGEM construct/vector used | No | ||||||||||||||||||||||||||
Plasmid/construct map | |||||||||||||||||||||||||||
Plasmid/construct sequence | |||||||||||||||||||||||||||
Restriction sites to linearize plasmid | |||||||||||||||||||||||||||
Selectable marker used to select the mutant parasite | hdhfr | ||||||||||||||||||||||||||
Promoter of the selectable marker | eef1a | ||||||||||||||||||||||||||
Selection (positive) procedure | pyrimethamine | ||||||||||||||||||||||||||
Selection (negative) procedure | No | ||||||||||||||||||||||||||
Additional remarks genetic modification | |||||||||||||||||||||||||||
Additional remarks selection procedure | |||||||||||||||||||||||||||
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Details of the target gene | |||||||||||||||||||||||||||
Gene Model of Rodent Parasite | PBANKA_0615200 | ||||||||||||||||||||||||||
Gene Model P. falciparum ortholog | PF3D7_0717500 | ||||||||||||||||||||||||||
Gene product | calcium-dependent protein kinase 4 | ||||||||||||||||||||||||||
Gene product: Alternative name | CDPK4 | ||||||||||||||||||||||||||
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Details of the genetic modification | |||||||||||||||||||||||||||
Name of the tag | c-myc | ||||||||||||||||||||||||||
Details of tagging | C-terminal | ||||||||||||||||||||||||||
Additional remarks: tagging | The mutated form of CDPK4 is tagged with 2xmyc | ||||||||||||||||||||||||||
Commercial source of tag-antibodies | |||||||||||||||||||||||||||
Type of plasmid/construct | (Linear) plasmid double cross-over | ||||||||||||||||||||||||||
PlasmoGEM (Sanger) construct/vector used | No | ||||||||||||||||||||||||||
Modified PlasmoGEM construct/vector used | No | ||||||||||||||||||||||||||
Plasmid/construct map | |||||||||||||||||||||||||||
Plasmid/construct sequence | |||||||||||||||||||||||||||
Restriction sites to linearize plasmid | |||||||||||||||||||||||||||
Selectable marker used to select the mutant parasite | hdhfr | ||||||||||||||||||||||||||
Promoter of the selectable marker | eef1a | ||||||||||||||||||||||||||
Selection (positive) procedure | pyrimethamine | ||||||||||||||||||||||||||
Selection (negative) procedure | No | ||||||||||||||||||||||||||
Additional remarks genetic modification | |||||||||||||||||||||||||||
Additional remarks selection procedure | |||||||||||||||||||||||||||
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