Back to search resultsSummaryRMgm-786
|
Successful modification | The parasite was generated by the genetic modification |
The mutant contains the following genetic modification(s) | Gene tagging |
Reference (PubMed-PMID number) |
Reference 1 (PMID number) : 22957089 |
MR4 number | |
top of page | |
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). |
top of page | |
The mutant parasite was generated by | |
Name PI/Researcher | N. Philip; A.P. Waters |
Name Group/Department | Wellcome Trust Centre for Molecular Parasitology, Institute of Infection, Immunity and Inflammation |
Name Institute | University of Glasgow |
City | Glasgow |
Country | UK |
top of page | |
Name of the mutant parasite | |
RMgm number | RMgm-786 |
Principal name | ppkl-gfp |
Alternative name | |
Standardized name | |
Is the mutant parasite cloned after genetic modification | Yes |
top of page | |
Phenotype | |
Asexual blood stage | Analyses of the mutant expressing GFP-tagged PPKLshows expression in schizonts, female gametocytes and ookinetes. |
Gametocyte/Gamete | Analyses of the mutant expressing GFP-tagged PPKLshows expression in schizonts, female gametocytes and ookinetes. Only female gametocytes and not males expressed PPKL. |
Fertilization and ookinete | Analyses of the mutant expressing GFP-tagged PPKL shows expression in schizonts, female gametocytes and ookinetes. Only female gametocytes and not males expressed PPKL. In ookinetes PPKL is localized predominantly in the cytoplasm with a particularly intense expression at the apical tip of the developing and mature ookinete. |
Oocyst | Not tested |
Sporozoite | Not tested |
Liver stage | Not tested |
Additional remarks phenotype | Mutant/mutation The mutant expresses a C-terminal GFP-tagged version of PPKL (protein phosphatase containing kelch-like domains). Protein (function) Phenotype analyses of mutants lacking expression of PPKL (RMgm-686, RMgm-687) indicate an essential role of PPKL in maturation of ookinetes. Phenotype Analyses of the mutant expressing GFP-tagged PPKLshows expression in schizonts, female gametocytes and ookinetes. Immunofluorescence studies using anti-PPKL polyclonal antibody showed expression in merozoites, female gametocytes, zygotes, ookinete, and sporozoites. Only female gametocytes and not males expressed PPKL. PPKL is localized predominantly in the cytoplasm with a particularly intense expression at the apical tip of the developing and mature ookinete. Additional information Other mutants |
top of page | |||||||||||||||||||||||||||
Details of the target gene | |||||||||||||||||||||||||||
Gene Model of Rodent Parasite | PBANKA_1329500 | ||||||||||||||||||||||||||
Gene Model P. falciparum ortholog | PF3D7_1466100 | ||||||||||||||||||||||||||
Gene product | protein phosphatase containing kelch-like domains | ||||||||||||||||||||||||||
Gene product: Alternative name | PPKL; PPalpha | ||||||||||||||||||||||||||
top of page | |||||||||||||||||||||||||||
Details of the genetic modification | |||||||||||||||||||||||||||
Name of the tag | GFP | ||||||||||||||||||||||||||
Details of tagging | C-terminal | ||||||||||||||||||||||||||
Additional remarks: tagging | |||||||||||||||||||||||||||
Commercial source of tag-antibodies | |||||||||||||||||||||||||||
Type of plasmid/construct | 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 | BglII | ||||||||||||||||||||||||||
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 | C-terminal GFP tagged PPKL parasite lines were generated using the PL0031 vector using single crossover homologous recombination containing 1 kb of the ppkl C-terminus amplified by using primers SacII-GU762 and BamHI-GU763 | ||||||||||||||||||||||||||
Additional remarks selection procedure | |||||||||||||||||||||||||||
| |||||||||||||||||||||||||||
top of page |