RMgmDB - Rodent Malaria genetically modified Parasites

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Summary

RMgm-1325
Malaria parasiteP. berghei
Genotype
TaggedGene model (rodent): PBANKA_0931200; Gene model (P.falciparum): PF3D7_1116800; Gene product: heat shock protein 101 | chaperone protein ClpB2 (HSP101)
Name tag: mCherry-3xMyc
Transgene
 Transgene not Plasmodium: GFP
Promoter: Gene model: PBANKA_0711900; Gene model (P.falciparum): PF3D7_0818900; Gene product: heat shock protein 70 (HSP70)
3'UTR: Gene model: Not available; Gene product: Not available
Insertion locus: Gene model: PBANKA_0931200; Gene product: heat shock protein 101 (HSP101)
Phenotype Asexual bloodstage; Oocyst; Sporozoite; Liver stage;
Last modified: 5 April 2016, 12:32
  *RMgm-1325
Successful modificationThe parasite was generated by the genetic modification
The mutant contains the following genetic modification(s) Gene tagging, 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-1325
Principal namehsp101-mCh(gfp)
Alternative name
Standardized name
Is the mutant parasite cloned after genetic modificationYes
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Phenotype
Asexual blood stageAbundant expression at the PVM in younger blood stages and in motile tubular structures in maturing trophozoites
Gametocyte/GameteNot tested
Fertilization and ookineteNot tested
OocystDistinct and abundant circumferential staining pattern in addition to uniform cytoplasmic distribution
SporozoiteAbundant apical staining in sporozoites
Liver stageNo expression until merozoite formation.
HSP101 expression was completely switched off during the first two days of intrahepatic growth.
Additional remarks phenotype

Mutant/mutation
The mutant expresses a mCherry tagged version of HSP101 and expresses GFP under the constitutive hsp70 promoter.

Protein (function)
Plasmodium parasites remodel their vertebrate host cells by translocating hundreds of proteins across an encasing membrane into the host cell cytosol via a putative export machinery termed PTEX (Plasmodium Translocon of EXported protein). HSP101 (PbANKA_094120), PTEX150 (PbANKA_100850), EXP2 (PbANKA_133430), PTEX88 (PbANKA_094130) and TRX2 (PbANKA_135800) have been identified as members of the PTEX complex.

Phenotype
Phenotype analyses indicate that HSP101 is located at the PVM of blood stages. In liver stages this protein is not detected in the PVM. HSP101 is also expressed in oocysts and sporozoites.

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
See link for other HSP101 mutants

  Tagged: Mutant parasite with a tagged gene
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Details of the target gene
Gene Model of Rodent Parasite PBANKA_0931200
Gene Model P. falciparum ortholog PF3D7_1116800
Gene productheat shock protein 101 | chaperone protein ClpB2
Gene product: Alternative nameHSP101
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Details of the genetic modification
Name of the tagmCherry-3xMyc
Details of taggingC-terminal
Additional remarks: tagging
Commercial source of tag-antibodies
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 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
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  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
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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 modification
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 Not available
Gene productNot available
Gene product: Alternative name
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 / InsertionInsertion locus
Gene Model of Parasite PBANKA_0931200
Gene productheat shock protein 101
Gene product: Alternative nameHSP101
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