RMgmDB - Rodent Malaria genetically modified Parasites

Summary

RMgm-4440
Malaria parasiteP. berghei
Genotype
TaggedGene model (rodent): PBANKA_1343800; Gene model (P.falciparum): PF3D7_1328800; Gene product: transcriptional regulatory protein sir2a (SIR2A)
Name tag: EGFP
Phenotype Asexual bloodstage; Gametocyte/Gamete; Fertilization and ookinete; Oocyst;
Last modified: 26 April 2018, 10:00
  *RMgm-4440
Successful modificationThe parasite was generated by the genetic modification
The mutant contains the following genetic modification(s) Gene tagging
Reference (PubMed-PMID number) Not published (yet)
MR4 number
Parent parasite used to introduce the genetic modification
Rodent Malaria ParasiteP. berghei
Parent strain/lineP. berghei ANKA
Name parent line/clone P. berghei ANKA cl15cy1
Other information parent lineA reference wild type clone from the ANKA strain of P. berghei (PubMed: PMID: 17406255).
The mutant parasite was generated by
Name PI/ResearcherReliga AA, Janse CJ, Waters AP
Name Group/DepartmentLeiden Malaria Research Group
Name InstituteLeiden University Medical Center
CityLeiden
CountryThe Netherlands
Name of the mutant parasite
RMgm numberRMgm-4440
Principal name503cl1
Alternative name
Standardized name
Is the mutant parasite cloned after genetic modificationYes
Phenotype
Asexual blood stageEvidence for PbSIR2A::GFP in asexual blood stages (mainly nuclear location).
Gametocyte/GameteEvidence for PbSIR2A::GFP in cytoplasm of gametocytes (higher in females than in males).
Fertilization and ookineteEvidence for PbSIR2A-GFP signal in mauture ookinetes concentrated peripherally near the apical end as well as the nucleus borders (circle-like staining). The characteristic apical-proximal localisation starts forming between 6 - 9h post-fertilisation and is most prominent in mature ookinetes, but is not present in young ookinetes (<6hrs post-fertilisation).
OocystEvidence for PbSIR2A::GFP in (cytoplasm of) oocysts.
SporozoiteNot tested
Liver stageNot tested
Additional remarks phenotype

Mutant/mutation
The mutant expresses a C-terminal GFP-tagged version of SIR2a

Protein (function)

Phenotype
Evidence for PbSIR2A::GFP in asexual blood stages (mainly nuclear location). Evidence for PbSIR2A::GFP in cytoplasm of gametocytes (higher in females than in males). Evidence for PbSIR2A-GFP signal in mature ookinetes concentrated peripherally near the apical end as well as the nucleus borders (circle-like staining). The characteristic apical-proximal localisation starts forming between 6 - 9h post-fertilisation and is most prominent in mature ookinetes, but is not present in young ookinetes (<6hrs post-fertilisation). Evidence for PbSIR2A::GFP in (cytoplasm of) oocysts.

See also mutant RMgm-4435 that lacks expression of SIR2a: Evidence is found that ookinetes do not traverse the midgut epithelium. No oocyst formation. No sporozoite formation.

Additional information

Other mutants
- see also mutant RMgm-4435 that lacks expression of SIR2a
- a mutant lacking sir2b - RMgm-4436
- a double gene knockout lacking both sir2b and sir2a (see mutant RMgm-4437).
 


  Tagged: Mutant parasite with a tagged gene
Details of the target gene
Gene Model of Rodent Parasite PBANKA_1343800
Gene Model P. falciparum ortholog PF3D7_1328800
Gene producttranscriptional regulatory protein sir2a
Gene product: Alternative nameSIR2A
Details of the genetic modification
Name of the tagEGFP
Details of taggingC-terminal
Additional remarks: tagging
Commercial source of tag-antibodies
Type of plasmid/construct(Linear) plasmid single cross-over
PlasmoGEM (Sanger) construct/vector usedNo
Modified PlasmoGEM construct/vector usedNo
Plasmid/construct map
Plasmid/construct sequence
Restriction sites to linearize plasmid
Selectable marker used to select the mutant parasitehdhfr
Promoter of the selectable markereef1a
Selection (positive) procedurepyrimethamine
Selection (negative) procedureNo
Additional remarks genetic modification
Additional remarks selection procedure
Primer information: Primers used for amplification of the target sequences  Click to view information
Primer information: Primers used for amplification of the target sequences  Click to hide information
Sequence Primer 1
Additional information primer 1
Sequence Primer 2
Additional information primer 2
Sequence Primer 3
Additional information primer 3
Sequence Primer 4
Additional information primer 4
Sequence Primer 5
Additional information primer 5
Sequence Primer 6
Additional information primer 6