RMgmDB - Rodent Malaria genetically modified Parasites

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Summary

RMgm-4643
Malaria parasiteP. berghei
Genotype
Transgene
 Transgene not Plasmodium: A fusion of YFP and Luciferase Firefly
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
Replacement locus: Gene model: Not available; Gene product: Not available (small subunit ribosomal rna gene (c-or d-type unit))
Phenotype Liver stage;
Last modified: 2 July 2019, 16:32
  *RMgm-4643
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) : 31189656
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/ResearcherBando H, Yamamoto M
Name Group/DepartmentDepartment of Immunoparasitology
Name InstituteOsaka University
CityOsaka
CountryJapan
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Name of the mutant parasite
RMgm numberRMgm-4643
Principal namePbYFPluc
Alternative name
Standardized name
Is the mutant parasite cloned after genetic modificationYes
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Phenotype
Asexual blood stageNot tested
Gametocyte/GameteNot tested
Fertilization and ookineteNot tested
OocystNot tested
SporozoiteNot tested
Liver stageExpression of YFP-luciferase in liver stages.
Additional remarks phenotype

Mutant/mutation
This mutant expresses the fusion protein YFP-luciferase under the control of the constitutive hsp70 promoter

Protein (function)

Phenotype
Expression of YFP-luciferase in liver stages (other stages not tested)

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: not Plasmodium
Transgene nameA fusion of YFP and Luciferase Firefly
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Details of the genetic modification
Inducable system usedNo
Additional remarks inducable system To express YFP and luciferase in P. berghei ANKA, construct encoding YFP and luciferase fusion protein (YFPluc) were cloned into the plasmid vector pSK-1-cssu (Niikura et al., 2013), which contains a selectable marker cassette encoding human dihydrofolate reductase (hdhfr) under control of the elongation factor 1 α (ef-1α) promoter, a GFP expression cassette under the control of the P. berghei heat shock protein 70-1 (hsp70-1) promoter, and homology arms of the cssu/dssu ribosomal gene locus of P. berghei. pSK-1-cssu was digested with NheI and BglII; therefore, GFP was removed. Subsequently, the YFPluc fragment was cloned into the NheI and BglII site of pSK-1-cssu. To obtain the P. berghei YFPluc, transfection was done by electroporation using the Amaxa nucleofector kit. The mutant parasites were selected by pyrimethamine for 2 wk, and then a clonal line expressing YFP and luciferase was obtained by limiting dilution.
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 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 / InsertionReplacement locus
Gene Model of Parasite Not available
Gene productNot available
Gene product: Alternative namesmall subunit ribosomal rna gene (c-or d-type unit)
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