RMgmDB - Rodent Malaria genetically modified Parasites

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Summary

RMgm-4441
Malaria parasiteP. berghei
Genotype
TaggedGene model (rodent): PBANKA_1213100; Gene model (P.falciparum): PF3D7_1014700; Gene product: prohibitin, putative (PHB2)
Name tag: mCherry-3xMyc
Transgene
 Transgene not Plasmodium: HSP70-3 fused to GFP
Promoter: Gene model: PBANKA_0914400; Gene model (P.falciparum): PF3D7_1134000; Gene product: heat shock protein 70 (HSP70-3, mtHSP70)
3'UTR: Gene model: PBANKA_0914400; Gene product: heat shock protein 70 (HSP70-3, mtHSP70)
Replacement locus: Gene model: PBANKA_1213100; Gene product: prohibitin, putative (PHB2)
Phenotype Asexual bloodstage; Gametocyte/Gamete; Fertilization and ookinete; Oocyst; Sporozoite; Liver stage;
Last modified: 27 April 2018, 17:34
  *RMgm-4441
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) : 29669282
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 Not applicable
Other information parent line
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The mutant parasite was generated by
Name PI/ResearcherMatz JM, Kooij TWA
Name Group/DepartmentDepartment of Molecular Parasitology, Institute of Biology
Name InstituteHumboldt University
CityBerlin
CountryGermany
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Name of the mutant parasite
RMgm numberRMgm-4441
Principal namePHB2-mCherry
Alternative name
Standardized name
Is the mutant parasite cloned after genetic modificationYes
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Phenotype
Asexual blood stagePHB2-mCherry expression with evidence for mitochondrial location
Gametocyte/GametePHB2-mCherry expression with evidence for mitochondrial location
Fertilization and ookinetePHB2-mCherry expression with evidence for mitochondrial location
OocystPHB2-mCherry expression with evidence for mitochondrial location
SporozoitePHB2-mCherry expression with evidence for mitochondrial location
Liver stagePHB2-mCherry expression with evidence for mitochondrial location
Additional remarks phenotype

Mutant/mutation
The mutant expresses a mCherry tagged version of PHB2 and expresses a GFP-tagged mitochondrial protein HSP70-3 (PBANKA_0914400)

Protein (function)
Proteins of the stomatin/prohibitin/flotillin/HfIK/C (SPFH) family are membrane-anchored and perform diverse cellular functions in different organelles. A systematic database search for Plasmodium SPFH proteins  uncovered four candidates that either display a close phylogenetic relationship to conserved members—prohibitin 1 (PHB1, PBANKA_0701500), prohibitin 2 (PHB2, PBANKA_1213100), and stomatin-like protein (STOML, PBANKA_0807100)- 0807100)—or fall into a unique sister group that contains related proteins from the phylum Myzozoa, which harbors the subphylum Apicomplexa. Accordingly,  the latter Plasmodium protein was named prohibitin-like protein (PHBL, PBANKA_0718600). All four proteins contain a central signature SPFH domain and a hydrophobic stretch Although the conventional Plasmodium SPFH proteins display a similarly high degree of conservation, PHBL sequences are rather divergent already among related Plasmodium species.

Phenotype
PHB2-mCherry expression in all stages with evidence for mitochondrial location

Additional information

Other mutants

  Tagged: Mutant parasite with a tagged gene
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Details of the target gene
Gene Model of Rodent Parasite PBANKA_1213100
Gene Model P. falciparum ortholog PF3D7_1014700
Gene productprohibitin, putative
Gene product: Alternative namePHB2
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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 nameHSP70-3 fused to GFP
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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 modificationAn mitochondrial marker cassette consisting of the promoter and N terminus of the mitochondrial gene HSP70-3 (PBANKA_0914400) fused to GFP
Additional remarks selection procedure
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Other details transgene
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Promoter
Gene Model of Parasite PBANKA_0914400
Gene Model P. falciparum ortholog PF3D7_1134000
Gene productheat shock protein 70
Gene product: Alternative nameHSP70-3, mtHSP70
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_0914400
Gene productheat shock protein 70
Gene product: Alternative nameHSP70-3, mtHSP70
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 / InsertionReplacement locus
Gene Model of Parasite PBANKA_1213100
Gene productprohibitin, putative
Gene product: Alternative namePHB2
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: StudioDrie StudioDrie; S. Mededovic and A. van Wigcheren