RMgmDB - Rodent Malaria genetically modified Parasites


Malaria parasiteP. berghei
MutatedGene model (rodent): PbANKA_1422900; Gene model (P.falciparum): PF3D7_0814600; Gene product: PIMMS22 protein, putative (Concavin)
Details mutation: P. berghei concavin (PIMMS22) gene replaced with P. falciparum Concavin gene (PF3D7_0814600)
PhenotypeNo phenotype has been described
Last modified: 16 April 2024, 12:53
Successful modificationThe 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) : 35403820
MR4 number
Parent parasite used to introduce the genetic modification
Rodent Malaria ParasiteP. berghei
Parent strain/lineP. berghei ANKA
Name parent line/clone RMgm-5127
Other information parent lineThe mutant lacks the Concavin (PIMMS22) gene. In addition, it does not contain a (positive) drug selectable marker. The selectable marker has been removed by negative selection.
The mutant parasite was generated by
Name PI/ResearcherKehrer J, Frischknecht F
Name Group/DepartmentIntegrative Parasitology, Center for Infectious Diseases
Name InstituteHeidelberg University Medical School
Name of the mutant parasite
RMgm numberRMgm-5128
Principal namePF3D7 complementation
Alternative name
Standardized name
Is the mutant parasite cloned after genetic modificationYes
Asexual blood stageNot tested
Gametocyte/GameteNot tested
Fertilization and ookineteNot tested
OocystNot different from wild type
SporozoiteNot different from wild type
Liver stageNot tested
Additional remarks phenotype

In the mutant the P. berghei concavin (PIMMS22) gene has been replaced with the P. falciparum Concavin (PIMMS22) gene (PF3D7_0814600). 

Protein (function)
See mutant RMgm-4999 for previous analyses of this protein (PIMMS22; Concavin):
PfPIMMS22 encodes a 393 amino acid (45 kDa) protein and PbPIMMS22 encodes a 393 aa (44 kDa) protein, both with no predicted signal peptide or transmembrane domain. The protein is highly conserved amongst Plasmodium orthologues. PyPIMMS22 (PY17X_1424900) was previously identified in salivary gland sporozoites through a  subtractive hybridization (SSH) profiling and termed sporozoite protein S15. The same protein was also identified in midgut oocyst sporozoites as an interacting partner to the apicomplexan specific RNA-binding protein, ALBA4, which is involved in mRNA regulation in gametocyte and midgut oocyst sporozoite development. PIMMS22 homologues are found in other apicomplexan parasites including Toxoplasma gondii, Neospora caninum and Eimeria with sequence identities to PIMMS22 ranging from 36% to 42%. InterPro domain analysis revealed no recognizable domain in PIMMS22. Evidence is presented for ookinete defects in invasion and traversal of the midgut epithelium (although this cannot completely explain the strongly reduced oocysts numbers, indicating reduced ookinete to oocyst transition after midgut traversal). Strongly reduced oocyst numbers. Strongly reduced sporozoite numbers. No infection of mice by the bite of infected mosquitoes.

From this study: Due to its impact on the convex-concave polarity of sporozoites we named the protein Concavin

No phenotypes of oocysts and sporozoites different from wild type were detected, indicating that P. falciparum concavin complements the function of P. berghei concavin

Additional information

Other mutants

  Mutated: Mutant parasite with a mutated gene
Details of the target gene
Gene Model of Rodent Parasite PbANKA_1422900
Gene Model P. falciparum ortholog PF3D7_0814600
Gene productPIMMS22 protein, putative
Gene product: Alternative nameConcavin
Details of the genetic modification
Short description of the mutationP. berghei concavin (PIMMS22) gene replaced with P. falciparum Concavin gene (PF3D7_0814600)
Inducable system usedNo
Short description of the conditional mutagenesisNot available
Additional remarks inducable system
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/yfcu
Promoter of the selectable markereef1a
Selection (positive) procedurepyrimethamine
Selection (negative) procedureNo
Additional remarks genetic modificationThe P. berghei 5`UTR was amplified using primers JK55 and JK179 and inserted into pL59 using KpnI and BstbI. Followed by the insertion of PF3D7_0814600 amplified with primers JK177 and JK178 from P. falciparum gDNA and digested with BstbI and NdeI. For transfection of concavin(-)NS parasites via double crossover the plasmid was digested with KpnI and SacII.
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