SummaryRMgm-4973
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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) : 34494883 |
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 cl15cy1 |
Other information parent line | A reference wild type clone from the ANKA strain of P. berghei (PubMed: PMID: 17406255). |
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The mutant parasite was generated by | |
Name PI/Researcher | Esveld SL, Kooij TWA, Huynen MA |
Name Group/Department | Center for Molecular and Biomolecular Informatics, Radboud Institute for Molecular Life Sciences |
Name Institute | Radboudumc |
City | Nijmegen |
Country | The Netherlands |
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Name of the mutant parasite | |
RMgm number | RMgm-4973 |
Principal name | 3xHA-PBANKA_0814400 |
Alternative name | |
Standardized name | |
Is the mutant parasite cloned after genetic modification | No |
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Phenotype | |
Asexual blood stage | Low expression of the tagged protein; no subcellular location could be assigned |
Gametocyte/Gamete | Not tested |
Fertilization and ookinete | Not tested |
Oocyst | Not tested |
Sporozoite | Not tested |
Liver stage | Not tested |
Additional remarks phenotype | Mutant/mutation Phenotype
Mitochondrial localization of seven out of 13 Plasmodium berghei orthologs (see below) was confirmed by epitope labeling and co-localization with a mitochondrial marker protein.
PBANKA_0715500 3xHA tag: mitochondrial
PBANKA_0109600 3xHA tag: mitochondrial
PBANKA_1024800 3xHA tag: mitochondrial
PBANKA_0310100 mOrange-3xHA tag: mitochondrial (3xHA tag; fixed blood stages)
PBANKA_0708900 3xHA tag: mitochondrial
PBANKA_1459800 3xHA tag: no subcellular location could be assigned
PBANKA_0814400 3xHA tag: no subcellular location could be assigned
PBANKA_1203200 3xHA-tagging: mitochondrial (3xHA tag; fixed blood stages)
PBANKA_0827900 3xHA tag: mitochondrial
PBANKA_1103500 3xHA tag: possible mitochondrial
PBANKA_0927100 3xHA tag: no subcellular location could be assigned
PBANKA_0914000 3xHA tag: possible mitochondrial
PBANKA_1350000 3xHA tag: possible mitochondrial
Additional information |
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Details of the target gene | |||||||||||||||||||||||||||
Gene Model of Rodent Parasite | PBANKA_0814400 | ||||||||||||||||||||||||||
Gene Model P. falciparum ortholog | PF3D7_0913400 | ||||||||||||||||||||||||||
Gene product | conserved protein, unknown function | ||||||||||||||||||||||||||
Gene product: Alternative name | |||||||||||||||||||||||||||
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Details of the genetic modification | |||||||||||||||||||||||||||
Name of the tag | triple-HA | ||||||||||||||||||||||||||
Details of tagging | C-terminal | ||||||||||||||||||||||||||
Additional remarks: tagging | |||||||||||||||||||||||||||
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 | unknown | ||||||||||||||||||||||||||
Selection (positive) procedure | pyrimethamine | ||||||||||||||||||||||||||
Selection (negative) procedure | No | ||||||||||||||||||||||||||
Additional remarks genetic modification | Tagging of target protein with a 3xHA tag (or combined fluorescent (mOrange) and epitope (3xHA) tag) and simultaneously introducing a mitochondrial targeted GFP marker by fusion of the promoter and N-terminus of HSP70-3 (PBANKA_0914400). | ||||||||||||||||||||||||||
Additional remarks selection procedure | |||||||||||||||||||||||||||
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Type and details of transgene | |||||||||||||||||||
Is the transgene Plasmodium derived | Transgene: not Plasmodium | ||||||||||||||||||
Transgene name | GFP | ||||||||||||||||||
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Details of the genetic modification | |||||||||||||||||||
Inducable system used | No | ||||||||||||||||||
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 | unknown | ||||||||||||||||||
Selection (positive) procedure | pyrimethamine | ||||||||||||||||||
Selection (negative) procedure | No | ||||||||||||||||||
Additional remarks genetic modification | |||||||||||||||||||
Additional remarks selection procedure | |||||||||||||||||||
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Other details transgene | |||||||||||||||||||
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Promoter | |||||||||||||||||||
Gene Model of Parasite | PBANKA_0914400 | ||||||||||||||||||
Gene Model P. falciparum ortholog | PF3D7_0831700 | ||||||||||||||||||
Gene product | heat shock protein 70 | ||||||||||||||||||
Gene product: Alternative name | HSP70x, HSP70-3, UIS24 | ||||||||||||||||||
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3'-UTR | |||||||||||||||||||
Gene Model of Parasite | PBANKA_0914400 | ||||||||||||||||||
Gene product | heat shock protein 70 | ||||||||||||||||||
Gene product: Alternative name | HSP70x, HSP70-3, UIS24 | ||||||||||||||||||
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Insertion/Replacement locus | |||||||||||||||||||
Replacement / Insertion | Insertion locus | ||||||||||||||||||
Gene Model of Parasite | PBANKA_0814400 | ||||||||||||||||||
Gene product | conserved protein, unknown function | ||||||||||||||||||
Gene product: Alternative name | |||||||||||||||||||
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