RMgmDB - Rodent Malaria genetically modified Parasites

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Summary

RMgm-1437
Malaria parasiteP. berghei
Genotype
Transgene
 Transgene Plasmodium: Gene model: Not available; Gene model (P.falciparum): PF3D7_0501100; Gene product: heat shock protein 40, type II (HSP40; PFE55)
Promoter: Gene model: PBANKA_1365500; Gene model (P.falciparum): Not available; Gene product: intra-erythrocytic P. berghei-induced structures protein 1 exported protein IBIS1 (Plasmodium exported protein IBIS1)
3'UTR: Gene model: PBANKA_1365500; Gene product: intra-erythrocytic P. berghei-induced structures protein 1 exported protein IBIS1 (Plasmodium exported protein IBIS1)
Replacement locus: Gene model: PBANKA_1365500; Gene product: intra-erythrocytic P. berghei-induced structures protein 1 exported protein IBIS1 (Plasmodium exported protein IBIS1)
Phenotype Asexual bloodstage;
Last modified: 16 April 2016, 20:24
  *RMgm-1437
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) : 27063072
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/ResearcherPetersen W; Przyborski JM
Name Group/DepartmentParasitology
Name InstitutePhilipps University Marburg
CityMarburg
CountryGermany
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Name of the mutant parasite
RMgm numberRMgm-1437
Principal namePFE55(GFP)
Alternative name
Standardized name
Is the mutant parasite cloned after genetic modificationNo
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Phenotype
Asexual blood stagePFE55(GFP)expressed in blood stages and exported into the cytoplasm of the infected red blood cell (diffuse staining pattern in cytoplasm)
Gametocyte/GameteNot tested
Fertilization and ookineteNot tested
OocystNot tested
SporozoiteNot tested
Liver stageNot tested
Additional remarks phenotype

Mutant/mutation
The mutant expresses a GFP tagged version of the P. falciparum-specific protein PFE55 (HSP40; PF3D7_0501100) under the control of the regulatory regions of the gene encoding the blood-stage exported protein IBIS1

Protein (function)
PFE55 (HSP40; PF3D7_0501100) is exported into the cytoplasm of P. falciparum-infected red blood cells and located in so-called J-dots (visualized as distinct fluorescent, punctate foci)

Phenotype
PFE55(GFP) expressed in blood stages and exported into the cytoplasm of the infected red blood cell (diffuse staining pattern in cytoplasm)

Additional information

Other mutants

  Transgene: Mutant parasite expressing a transgene
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Type and details of transgene
Is the transgene Plasmodium derived Transgene: Plasmodium
Gene Model of Parasite Not available
Gene Model P. falciparum ortholog PF3D7_0501100
Gene productheat shock protein 40, type II
Gene product: Alternative nameHSP40; PFE55
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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
Promoter of the selectable markereef1a
Selection (positive) procedurepyrimethamine
Selection (negative) procedureNo
Additional remarks genetic modificationFor expression in Plasmodium berghei, HSP40-GFP was amplified from the pARL2 construct and flanked by the 50 and 30 regions of IBIS1, amplified from P. berghei ANKA strain genomic DNA, in construct b3D.DT^H.^D containing the selectable marker human dihydrofolate reductase (hDHFR). Prior to transfection, the vector was linearised by PacI digestion to stimulate integration within the IBIS1 5' region
Additional remarks selection procedure
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Other details transgene
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Promoter
Gene Model of Parasite PBANKA_1365500
Gene Model P. falciparum ortholog Not available
Gene productintra-erythrocytic P. berghei-induced structures protein 1 exported protein IBIS1
Gene product: Alternative namePlasmodium exported protein IBIS1
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_1365500
Gene productintra-erythrocytic P. berghei-induced structures protein 1 exported protein IBIS1
Gene product: Alternative namePlasmodium exported protein IBIS1
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_1365500
Gene productintra-erythrocytic P. berghei-induced structures protein 1 exported protein IBIS1
Gene product: Alternative namePlasmodium exported protein IBIS1
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