SummaryRMgm-1501
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Successful modification | The parasite was generated by the genetic modification |
The mutant contains the following genetic modification(s) | Gene mutation |
Reference (PubMed-PMID number) |
Reference 1 (PMID number) : 27427910 |
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-1500 |
Other information parent line | This is a P. berghei mutant lacking the pat gene (PBANKA_0303900) |
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The mutant parasite was generated by | |
Name PI/Researcher | Kehrer J; Mair GR |
Name Group/Department | Integrative Parasitology, Center for Infectious Diseases |
Name Institute | University of Heidelberg Medical School |
City | Heidelberg |
Country | Germany |
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Name of the mutant parasite | |
RMgm number | RMgm-1501 |
Principal name | Δpat;patPF3D7::mcherry |
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 | Not different from wild type |
Fertilization and ookinete | Not tested |
Oocyst | Not different from wild type |
Sporozoite | Not different from wild type |
Liver stage | Not tested |
Additional remarks phenotype | Mutant/mutation A mutant lacking expression of PAT (Δpat; RMgm-1500); A mutant in which the endogeneous pat gene is replaced with the pat gene of P. falciparum 3D7: Δpat;pat(PF3D7)::mcherry; RMgm-1501) A mutant expressing GFP-tagged PAT and mCherry-tagged G377 (pat::gfp;g377:mcherry; RMgm-1503): A mutant lacking expression of PAT and expressing mCherry-tagged G377 (PBANKA_1463000) and a mutant lacking expression of PAT and expressing mCherry-tagged PPLP2 (PBANKA_1463000) A mutant expressing a GFP-tagged version of TRAP (ss::gfp::trap; RMgm-1505) A 'promoter-swap' mutant expressing PAT under the promoter of the ccp1/lap2 promoter (PBANKA_1300700)(patccp.PP; RMgm-1506) |
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Details of the target gene | |||||||||||||||||||||||||||
Gene Model of Rodent Parasite | PBANKA_0303900 | ||||||||||||||||||||||||||
Gene Model P. falciparum ortholog | PF3D7_0206200 | ||||||||||||||||||||||||||
Gene product | pantothenate transporter | ||||||||||||||||||||||||||
Gene product: Alternative name | PAT | ||||||||||||||||||||||||||
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Details of the genetic modification | |||||||||||||||||||||||||||
Short description of the mutation | The endogenous pat gene has been replaced with the P. falciparum 3D7 pat gene | ||||||||||||||||||||||||||
Inducable system used | No | ||||||||||||||||||||||||||
Short description of the conditional mutagenesis | Not available | ||||||||||||||||||||||||||
Additional remarks inducable system | |||||||||||||||||||||||||||
Type of plasmid/construct | (Linear) plasmid single 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 | WR99210 | ||||||||||||||||||||||||||
Selection (negative) procedure | No | ||||||||||||||||||||||||||
Additional remarks genetic modification | A 1704 bp fragment encompassing the near-complete ORF of P. falciparum PAT PF3D7_0206200 (starting from amino acid 4) was PCR-amplified from P. falciparum PF3D7 genomic DNA with primers g3110 (BsmI) and g3111 (BamHI) and cloned into pLIS0201 resulting in an almost complete exchange of P. berghei pat for P. falciparum PF3D7_0206200; only the first three amino acids of the fusion gene remain from P. berghei (MNA), the remainder is P. falciparum sequence. The final plasmid pLIS0301 was digested with EcoRV, precipitated with 2.5 volumes ethanol and transfected into Δpat schizonts. | ||||||||||||||||||||||||||
Additional remarks selection procedure | |||||||||||||||||||||||||||
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