RMgmDB - Rodent Malaria genetically modified Parasites

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Summary

RMgm-4131
Malaria parasiteP. berghei
Genotype
TaggedGene model (rodent): PBANKA_1032100; Gene model (P.falciparum): PF3D7_1410400; Gene product: rhoptry-associated protein 1 (RAP1)
Name tag: triple-HA
Phenotype Asexual bloodstage;
Last modified: 15 April 2017, 14:35
  *RMgm-4131
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) : 28205319
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/ResearcherGhosh S; de Koning-Ward TF
Name Group/DepartmentSchool of Medicine
Name InstituteDeakin University
CityWaurn Ponds
CountryAustralia
Name of the mutant parasite
RMgm numberRMgm-4131
Principal nameRAP1-HA
Alternative name
Standardized name
Is the mutant parasite cloned after genetic modificationNo
Phenotype
Asexual blood stageRAP1-HA expression revealed by Western analysis and IFA using anti-HA antibodies (punctate staining pattern in schizonts)
Gametocyte/GameteNot tested
Fertilization and ookineteNot tested
OocystNot tested
SporozoiteNot tested
Liver stageNot tested
Additional remarks phenotype

Mutant/mutation
The mutant expresses a C-terminal (triple) HA and and (single) streptavidin-tagged version of RAP1

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

RAP1-HA expression revealed by Western analysis and IFA using anti-HA antibodies (punctate staining pattern in schizonts)

Additional information
Unsuccessful attempts to disrupt P. berghei rap1 and rap2/3 indicate an essential function during blood stage growth/development/invasion. Conditional knock-down of P. berghei rap1 (RMgm-4132) and rap2/3 (RMgm-4133) provided evidence for a role of these proteins the formation of the parasitophorous vacuole membrane (PVM) (and not in invasion of red blood cells) 

Other mutants
See RAP1 and RAP2/3


  Tagged: Mutant parasite with a tagged gene
Details of the target gene
Gene Model of Rodent Parasite PBANKA_1032100
Gene Model P. falciparum ortholog PF3D7_1410400
Gene productrhoptry-associated protein 1
Gene product: Alternative nameRAP1
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 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 parasitetgdhfr
Promoter of the selectable markerpbdhfr
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