RMgmDB - Rodent Malaria genetically modified Parasites

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Summary

RMgm-4438
Malaria parasiteP. berghei
Genotype
TaggedGene model (rodent): PBANKA_0924800; Gene model (P.falciparum): PF3D7_1123500; Gene product: golgi protein 2 (GP2)
Name tag: GFP
Phenotype Asexual bloodstage; Gametocyte/Gamete; Fertilization and ookinete; Oocyst;
Last modified: 20 April 2018, 15:18
  *RMgm-4438
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) : 29579782
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/ResearcherHallée S, Mair GR, Richard D
Name Group/DepartmentCentre de recherche en infectiologie
Name InstituteCHU de Québec-Université Laval
CityQuebec City, Quebec
CountryCanada
Name of the mutant parasite
RMgm numberRMgm-4438
Principal namegp2::gfp
Alternative name
Standardized name
Is the mutant parasite cloned after genetic modificationYes
Phenotype
Asexual blood stageexpression in blood stages
Gametocyte/Gameteexpression in gametocytes
Fertilization and ookineteexpression in ookinetes
Oocystexpression in oocysts
SporozoiteNot tested
Liver stageNot tested
Additional remarks phenotype

Mutant/mutation
The mutant expresses a C-terminal GFP-tagged version of GP2

Protein (function)
Eveidence is provided that GP2 (Golgi protein 2) forms a complex with Golgi protein 1 (PF3D7_1320000; rhoptry protein 2, putative; golgi protein 1; PBANKA_1418300). The Golgi Protein complex localizes to the cis-Golgi throughout the erythrocytic cycle and  potentially  also  during  the  mosquito  stages.

Phenotype
Expression of GFPtagged GP2 in multiple blood and mosquito stages

Additional information
The  extensive  colocalization  between  GP2  and  the  cis-Golgi apparatus  marker  Endoplasmic  reticulum  Retention  Defective  2  (ERD2)  confirmed that GP2 resides in the Golgi throughout the blood stages. This staining pattern was conserved in the rodent malaria parasite P. berghei, where we tagged the endogenous locus of the orthologous gene PBANKA_092480 with GFP. The tagged protein was  also  detected in  gametocytes,  as  well  as  all  mosquito  stage  parasites  including  the ookinete, midgut oocysts, and sporozoites providing evidence for the constitutive expression of GP2 throughout the life cycle of the parasite. In addition, the staining pattern of PbGP2 in the mosquito stages resembled what was previously described with PbGP1 which suggests that they might perhaps also form a complex in this phase of the life cycle

Other mutants
RMgm-4439: a mutant lacking GP2


  Tagged: Mutant parasite with a tagged gene
Details of the target gene
Gene Model of Rodent Parasite PBANKA_0924800
Gene Model P. falciparum ortholog PF3D7_1123500
Gene productgolgi protein 2
Gene product: Alternative nameGP2
Details of the genetic modification
Name of the tagGFP
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 parasiteunknown
Promoter of the selectable markerunknown
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