RMgmDB - Rodent Malaria genetically modified Parasites

Summary

RMgm-1332
Malaria parasiteP. berghei
Genotype
Transgene
 Transgene not Plasmodium: GFP fused to the signal peptide and ER-retention sequences of PBANKA_081890
Promoter: Gene model: PBANKA_0711900; Gene model (P.falciparum): PF3D7_0818900; Gene product: heat shock protein 70 (HSP70)
3'UTR: Gene model: PBANKA_1340000; Gene product: dihydrofolate synthase/folylpolyglutamate synthase, putative (PbDHFS-FPGS)
Phenotype Asexual bloodstage;
Last modified: 3 October 2015, 18:53
  *RMgm-1332
Successful modificationThe parasite was generated by the genetic modification
The mutant contains the following genetic modification(s) Introduction of a transgene
Reference (PubMed-PMID number) Reference 1 (PMID number) : 26219962
MR4 number
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Parent parasite used to introduce the genetic modification
Rodent Malaria ParasiteP. berghei
Parent strain/lineP. berghei ANKA
Name parent line/clone P. berghei ANKA cl15cy1
Other information parent lineA reference wild type clone from the ANKA strain of P. berghei (PubMed: PMID: 17406255).
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The mutant parasite was generated by
Name PI/ResearcherMatz, JM; Kooij, TW
Name Group/DepartmentMedical Microbiology
Name InstituteRadboudumc
CityNijmegen
CountryThe Netherlands
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Name of the mutant parasite
RMgm numberRMgm-1332
Principal nameGFPer
Alternative name
Standardized name
Is the mutant parasite cloned after genetic modificationYes
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Phenotype
Asexual blood stageStrong GFP-staining of the endoplasmic reticulum (ER)
Gametocyte/GameteNot tested
Fertilization and ookineteNot tested
OocystNot tested
SporozoiteNot tested
Liver stageNot tested
Additional remarks phenotype

Mutant/mutation
The mutant expresses GFP marker fused to the signal peptide and ER-retention sequences of PBANKA_081890 (heat shock protein 70, putative; HSP70-2, BIP), which labels the parasite’s ER. This reporter transgene is expressed under the control of the strong and constitutive hsp70 promoter.

Protein (function)
 
Phenotype
Strong GFP-staining of the endoplasmic reticulum (ER) in blood stages

Additional information
In the paper a mutant is described that expresses mCherry-tagged HSP101 and expresses GFP in the parasitophorous vacuole (PV). This mutant is obtained by (mosquito) crossing of mutant RMgm-1327 with mutant RMgm-1333

In the paper a mutant is described that expresses mCherry-tagged HSP101 and expresses GFP in the endoplasmatic reticulum (ER). This mutant is obtained by (mosquito) crossing of mutant RMgm-1327 with mutant RMgm-1332

Other mutants
RMgm-1333: A mutant expressing a GFP marker fused to the signal peptide (WITHOUT ER-retention sequences) of PBANKA_081890 (heat shock protein 70, putative; HSP70-2, BIP), which labels the parasite’s parasitophorous vacuole (PV). This reporter transgene is expressed under the control of the strong and constitutive hsp70 promoter.

  Transgene: Mutant parasite expressing a transgene
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Type and details of transgene
Is the transgene Plasmodium derived Transgene: not Plasmodium
Transgene nameGFP fused to the signal peptide and ER-retention sequences of PBANKA_081890
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Details of the genetic modification
Inducable system usedNo
Additional remarks inducable system
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 markerpbdhfr
Selection (positive) procedurepyrimethamine
Selection (negative) procedureNo
Additional remarks genetic modificationThe pBAT-SIL6 plasmid (see RMgm-757) was used to generate this mutant.
The linearized plasmid was integrated by double cross-over integration in a locus on P. berghei chromosome 6 between two hypothetical open reading frames (PBANKA_061210 and PBANKA_061220), both displaying only limited gene expression in the stages studied thus far. The integration of pBAT-SIL6 at this locus was designed to occur 1 kb 3' of PBANKA_061210 and 0.9 kb of PBANKA_260 061220.
Additional remarks selection procedure
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Other details transgene
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Promoter
Gene Model of Parasite PBANKA_0711900
Gene Model P. falciparum ortholog PF3D7_0818900
Gene productheat shock protein 70
Gene product: Alternative nameHSP70
Primer information details of the primers used for amplification of the promoter sequence  Click to view information
Primer information details of the primers used for amplification of the promoter sequence  Click to hide information
Sequence Primer 1
Additional information primer 1
Sequence Primer 2
Additional information primer 2
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3'-UTR
Gene Model of Parasite PBANKA_1340000
Gene productdihydrofolate synthase/folylpolyglutamate synthase, putative
Gene product: Alternative namePbDHFS-FPGS
Primer information details of the primers used for amplification the 3'-UTR sequences  Click to view information
Primer information details of the primers used for amplification the 3'-UTR sequences  Click to hide information
Sequence Primer 1
Additional information primer 1
Sequence Primer 2
Additional information primer 2
Insertion/Replacement locus
Replacement / InsertionNot available
Gene Model of Parasite Not available
Gene productNot available
Gene product: Alternative name
Primer information details of the primers used for amplification of the target sequences  Click to view information
Primer information details of the 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
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Conceptual design of the database: Dr. C.J. Janse (Leiden Malaria Research Group, Leiden, The Netherlands).
Technical design and realization: S. Mededovic and A. van Wigcheren