SummaryRMgm-12
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*RMgm-12| 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 |
Disrupted: Mutant parasite with a disrupted gene| top of page | |||||||||||||||||||||||||
| 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 | |||||||||||||||||||||||||
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Primer information: Primers used for amplification of the target sequences
![]() Primer information: Primers used for amplification of the target sequences
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