RMgmDB - Rodent Malaria genetically modified Parasites

Summary

RMgm-5214
Malaria parasiteP. berghei
Genotype
TaggedGene model (rodent): PBANKA_1457700; Gene model (P.falciparum): PF3D7_1244500; Gene product: PIMMS57 protein (PSOP26)
Name tag: triple-HA
Phenotype Gametocyte/Gamete; Fertilization and ookinete;
Last modified: 20 June 2022, 14:25
  *RMgm-5214
Successful modificationThe 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) : 35606790
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 2.34
Other information parent lineP. berghei ANKA 2.34 is a cloned, gametocyte producer line of the ANKA strain (PubMed: PMID: 15137943).
The mutant parasite was generated by
Name PI/ResearcherWang PP, Cao Y
Name Group/DepartmentDepartment of Immunology, College of Basic Medical Sciences
Name InstituteChina Medical University
CityShenyang
CountryChina
Name of the mutant parasite
RMgm numberRMgm-5214
Principal namePSOP26::HA
Alternative name
Standardized name
Is the mutant parasite cloned after genetic modificationYes
Phenotype
Asexual blood stageNot different from wild type
Gametocyte/Gameteevidence for localization of the protein at the membrane of male and female gametes (absent in gametocytes before activation) and at the membrane of ookinetes.
Fertilization and ookineteevidence for localization of the protein at the membrane of male and female gametes (absent in gametocytes before activation) and at the membrane of ookinetes.
OocystNot tested
SporozoiteNot tested
Liver stageNot tested
Additional remarks phenotype

Mutant/mutation
The mutant expresses a C-terminal 3xHA tagged version of PSOP26 (PIMMS57 protein)

Protein (function)
See also mutant RMgm-4998

Phenotype
Evidence for localization of the protein at the membrane of male and female gametes (absent in gametocytes before activation) and at the membrane of ookinetes.

See mutant RMgm-5213 that lacks expression of PSOP26 with the following phenotype: A strong reduction (>69%) in mature ookinete numbers. Reduced oocyst numbers (80% reduction). 

See also mutant RMgm-4998 lacking expression of PSOP26/PIMMS57 protein where they found a (slightly) different phenotype ('wild type-like ookinete production but reduced oocyst numbers').

Additional information
See also mutant RMgm-5001 where the found evidence for expression in (the membrane of) ookinetes of PSOP26/PIMMS57 

Other mutants


  Tagged: Mutant parasite with a tagged gene
Details of the target gene
Gene Model of Rodent Parasite PBANKA_1457700
Gene Model P. falciparum ortholog PF3D7_1244500
Gene productPIMMS57 protein
Gene product: Alternative namePSOP26
Details of the genetic modification
Name of the tagtriple-HA
Details of taggingC-terminal
Additional remarks: tagging
Commercial source of tag-antibodies
Type of plasmid/construct(Linear) plasmid double 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/yfcu
Promoter of the selectable markereef1a
Selection (positive) procedurepyrimethamine
Selection (negative) procedureNo
Additional remarks genetic modificationTo tag the endogenous psop26 with a 3 × HA tag, a 769-basepair (bp) fragment [nucleotide (nt) positions 1554–2322 bp] of the psop26 gene was amplified from P. berghei genomic DNA gDNA and ligated into the ApaI and SacII sites of the pL0034 plasmid as the 5′ homologous region (5R). Then, 526 bp of the psop26 3′ untranslated region (UTR) (nt + 1– + 526 bp) was amplified and inserted between the KpnI and EcoRI sites as the 3′ homologous region (3R), to yield the final plasmid pL0034-PSOP26-3 × HA.
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