RMgmDB - Rodent Malaria genetically modified Parasites

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Summary

RMgm-961
Malaria parasiteP. berghei
Genotype
TaggedGene model (rodent): PBANKA_0416500; Gene model (P.falciparum): PF3D7_0904900; Gene product: copper-transporting ATPase (CuTP)
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: PBANKA_1340000; Gene product: dihydrofolate synthase/folylpolyglutamate synthase, putative (Dhfs-fpgs)
Replacement locus: Gene model: PBANKA_0416500; Gene product: copper-transporting ATPase (CuTP)
Phenotype Asexual bloodstage; Gametocyte/Gamete; Fertilization and ookinete; Sporozoite; Liver stage;
Last modified: 27 July 2015, 18:45
  *RMgm-961
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) : 24237419
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/ResearcherS, Kenthirapalan; T.W. Kooij
Name Group/DepartmentParasitology Unit
Name InstituteMax Planck Institute for Infection Biology
CityBerlin
CountryGermany
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Name of the mutant parasite
RMgm numberRMgm-961
Principal namecutp::tag
Alternative name
Standardized name
Is the mutant parasite cloned after genetic modificationYes
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Phenotype
Asexual blood stageExpression of the mCherry-3xMyc-tagged version of CuTP(punctate location in cytoplasm)
Gametocyte/GameteExpression of the mCherry-3xMyc-tagged version of CuTP(punctate location in cytoplasm)
Fertilization and ookineteExpression of the mCherry-3xMyc-tagged version of CuTP in ookinetes(punctate location in cytoplasm)
OocystNot tested
SporozoiteExpression of the mCherry-3xMyc-tagged version of CuTP(punctate location in cytoplasm)
Liver stageExpression of the mCherry-3xMyc-tagged version of CuTP(diffuse staining of cytoplasm)
Additional remarks phenotype

Mutant/mutation
The mutant expresses a C-terminal  mCherry-3xMyc-tagged version of PBANKA_041650 (copper-transporting ATPase, putative; CuTP) and GFP under control of the constitutiive hsp70 promoter.

Protein (function)
Copper serves as a cofactor in metalloenzymes and catalyzes electron transfer reactions as well as the generation of potentially toxic reactive oxygen species.
Plasmodium species harbor at least three genes, which are predicted to be involved in copper transport and the maintenance of copper homeostasis: two proteins harboring a putative Ctr copper transporter domain (Pfam04145; PBANKA_102150 and PBANKA_130290) and one copper-transporting P-type ATPase (CuTP; PBANKA_041650).
Phenotype analyses of a mutant lacking CuTP (RMgm-960) indicate a strongly reduced fertility of male and female gametocytes/gametes of parasites lacking PBANKA_041650 (copper-transporting ATPase, putative) . The protein has a dispensable role during asexual blood stage growth/multiplication.

Phenotype
Phenotype analyses of a mutant lacking CuTP (RMgm-960) indicate a strongly reduced fertility of male and female gametocytes/gametes of parasites lacking PBANKA_041650 (copper-transporting ATPase, putative) . The protein has a dispensable role during asexual blood stage growth/multiplication.

Analyses of the mutant expressing a a C-terminal mCherry-3xMyc-tagged version of CuTP demonstrates that CuTP is predominantly located in vesicular bodies in the cytoplasm of the parasite.

Additional information

Other mutants
RMgm-960: a mutant lacking expression of PBANKA_041650 (copper-transporting ATPase, putative; CuTP)

  Tagged: Mutant parasite with a tagged gene
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Details of the target gene
Gene Model of Rodent Parasite PBANKA_0416500
Gene Model P. falciparum ortholog PF3D7_0904900
Gene productcopper-transporting ATPase
Gene product: Alternative nameCuTP
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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 modificationAn intermediate pBAT-derived vector was used (see RMgm-757)
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 modificationAn intermediate pBAT-derived vector was used (see RMgm-757)
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 nameDhfs-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 / InsertionReplacement locus
Gene Model of Parasite PBANKA_0416500
Gene productcopper-transporting ATPase
Gene product: Alternative nameCuTP
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