SummaryRMgm-12
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Successful modification | The parasite was generated by the genetic modification |
The mutant contains the following genetic modification(s) | Gene disruption |
Reference (PubMed-PMID number) |
Reference 1 (PMID number) : 15137943 |
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 | O. Billker |
Name Group/Department | Department of Biological Sciences |
Name Institute | Imperial College London |
City | London |
Country | United Kingdom |
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Name of the mutant parasite | |
RMgm number | RMgm-12 |
Principal name | ∆CDPK4 |
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 male gametes are formed; Male gametocytes are not affected in the ability to 'round up'and lyse the host cell; in male gametocytes axoneme formation, formation of mitotic spindles and DNA synthesis is inhibited/blocked. Female gametes are fertile; mutant female gametes fertilised by wild type male gametes produce ookinetes. However, these ookinetes are strongly affected in their ability to produce oocysts |
Fertilization and ookinete | See the description of the phenotype of Gametocytes/Gametes and Additional remarks phenotype |
Oocyst | See the description of the phenotype of Gametocytes/Gametes and 'Additional remarks phenotype'. |
Sporozoite | Not tested |
Liver stage | Not tested |
Additional remarks phenotype | Mutant/mutation Protein (function) Phenotype Additional information Other mutants |
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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 | |||||||||||||||||||||||||
Inducable system used | No | ||||||||||||||||||||||||
Additional remarks inducable system | |||||||||||||||||||||||||
Type of plasmid/construct used | 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 | |||||||||||||||||||||||||
Partial or complete disruption of the gene | Complete | ||||||||||||||||||||||||
Additional remarks partial/complete disruption | |||||||||||||||||||||||||
Selectable marker used to select the mutant parasite | tgdhfr | ||||||||||||||||||||||||
Promoter of the selectable marker | pbdhfr | ||||||||||||||||||||||||
Selection (positive) procedure | pyrimethamine | ||||||||||||||||||||||||
Selection (negative) procedure | No | ||||||||||||||||||||||||
Additional remarks genetic modification | The ∆CDPK4 mutant has been complemented with an intact copy of the cdpk4 gene, tagged with 2 copies of a carboxy-terminal c-myc epitope tag (insertion vector). Complementation resulted in the restoration of the wild type phenotype (wildtype male gamete formation, fertilisation and ookinete infectivity). | ||||||||||||||||||||||||
Additional remarks selection procedure | |||||||||||||||||||||||||
Primer information: Primers used for amplification of the target sequences
Primer information: Primers used for amplification of the target sequences
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