Summary

RMgm-4428
Malaria parasiteP. berghei
Genotype
DisruptedGene model (rodent): PBANKA_1101400; Gene model (P.falciparum): PF3D7_0501600; Gene product: rhoptry-associated protein 2 (rhoptry-associated protein 2/3; RAP2/3)
Phenotype Asexual bloodstage;
Last modified: 9 April 2018, 13:46
  *RMgm-4428
Successful modificationThe parasite was generated by the genetic modification
The mutant contains the following genetic modification(s) Gene disruption
Reference (PubMed-PMID number) Reference 1 (PMID number) : 29307564
MR4 number
Parent parasite used to introduce the genetic modification
Rodent Malaria ParasiteP. berghei
Parent strain/lineP. berghei ANKA
Name parent line/clone Not applicable
Other information parent line
The mutant parasite was generated by
Name PI/ResearcherNiikura M, Kobayashi F
Name Group/DepartmentDepartment of Infectious Diseases
Name InstituteKyorin University School of Medicine
CityTokyo
CountryJapan
Name of the mutant parasite
RMgm numberRMgm-4428
Principal nameΔrap2/3
Alternative name
Standardized name
Is the mutant parasite cloned after genetic modificationYes
Phenotype
Asexual blood stageParasitemia in Δrap2/3 parasite-infected mice was similar to that in control-parasite-infected mice until day 7 p.i. About 50% of B6 mice infected with Δrap2/3 parasites showed cerebral complications and died in a similar manner to mice infected with Pb ANKA at days 7-10 p.i., but approximately 40% of the mice ultimately cleared the parasites by day 60 p.i.
Gametocyte/GameteNot tested
Fertilization and ookineteNot tested
OocystNot tested
SporozoiteNot tested
Liver stageNot tested
Additional remarks phenotype

Mutant/mutation
The mutant lacks expression of RAP2/3 (and expresses GFP under control of the hsp70 promoter)

Protein (function)
In P. falciparum a low-molecular-weight protein complex, consisting of three non-covalently linked members, RAP1, RAP2 and RAP3 localises to the rhoptries (rhoptry bulb) of merozoites. RAP2 and RAP3 are encoded by adjacent genes, organised in a head to tail fashion. Both rap2 and rap3 show a high homology with the single copy, syntenic gene, rap2/3 of P. berghei.

Phenotype
Parasitemia in Δrap2/3 parasite-infected mice was similar to that in control-parasite-infected mice until day 7 p.i. About 50% of B6 mice infected with Δrap2/3 parasites showed cerebral complications and died in a similar manner to mice infected with Pb ANKA at days 7-10 p.i., but approximately 40% of the mice ultimately cleared the parasites by day 60 p.i.

Additional information

Other mutants

See link PBANKA_1101400  for other rap2/3 mutants (unsuccessful attempts to disrupt the rap2/3 gene and rap2/3 knock-out mutants with slow growing asexual blood stage phenotype)


  Disrupted: Mutant parasite with a disrupted gene
Details of the target gene
Gene Model of Rodent Parasite PBANKA_1101400
Gene Model P. falciparum ortholog PF3D7_0501600
Gene productrhoptry-associated protein 2
Gene product: Alternative namerhoptry-associated protein 2/3; RAP2/3
Details of the genetic modification
Inducable system usedNo
Additional remarks inducable system
Type of plasmid/construct used(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
Partial or complete disruption of the geneComplete
Additional remarks partial/complete disruption
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