RMgmDB - Rodent Malaria genetically modified Parasites

Summary

RMgm-4976
Malaria parasiteP. berghei
Genotype
DisruptedGene model (rodent): PBANKA_1004000; Gene model (P.falciparum): PF3D7_0406400; Gene product: cytosolic glyoxalase II (cGLO2)
PhenotypeNo phenotype has been described
Last modified: 10 May 2021, 18:41
  *RMgm-4976
Successful modificationThe parasite was generated by the genetic modification
The mutant contains the following genetic modification(s) Gene disruption
Reference (PubMed-PMID number) Reference 1 (PMID number) : 33667710
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/ResearcherSoga A, Fukumoto S
Name Group/DepartmentNational Research Center for Protozoan Diseases
Name InstituteObihiro University of Agriculture and Veterinary Medicine
CityInada-cho, Obihiro, Hokkaido
CountryJapan
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Name of the mutant parasite
RMgm numberRMgm-4976
Principal namecgloII KO
Alternative name
Standardized name
Is the mutant parasite cloned after genetic modificationYes
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Phenotype
Asexual blood stageNot different from wild type
Gametocyte/GameteNot tested
Fertilization and ookineteNot tested
OocystNot tested
SporozoiteNot tested
Liver stageNot different from wild type
Additional remarks phenotype

Mutant/mutation
The mutant lacks expression of cGLO2

Protein (function)
The malaria parasite also possesses a glyoxalase system in both the cytosol and apicoplast. The cytosolic pathway consists of GloI and cytosolic GloII (cGloII), and the apicoplast pathway consists of a targeted gloII (tGloII).

Phenotype
Only asexual blood stage development was analysed and the prepatent period in mice infected intravenously with 10.000 sporozoites. Liver stage development in cultured hepatocytes was determined by real-time RT-PCR. No phenotype was detected
 
Additional information
In this study both tGLO2 (targeted glyoxalase II: PF3D7_1205700) and cGLO2 (cytosolic glyoxalase II: PF3D7_0406400; RMgm-4976) were deleted. No phenotype was detected in blood and liver stages of both deletion mutants. However, evidence is presented that a 'double knock-out mutant' lacking both genes (RMgm-4977) shows a delayed liver stage development. 
 
Other mutants

  Disrupted: Mutant parasite with a disrupted gene
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Details of the target gene
Gene Model of Rodent Parasite PBANKA_1004000
Gene Model P. falciparum ortholog PF3D7_0406400
Gene productcytosolic glyoxalase II
Gene product: Alternative namecGLO2
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Details of the genetic modification
Inducable system usedNo
Additional remarks inducable system
Type of plasmid/construct used(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
Partial or complete disruption of the geneComplete
Additional remarks partial/complete disruption
Selectable marker used to select the mutant parasiteblasticidin S deaminase (bsd)
Promoter of the selectable markerhsp70
Selection (positive) procedureblasticydin in vitro
Selection (negative) procedureNo
Additional remarks genetic modificationsee mutant RMgm-4180 and RMgm-4455 for the transfection and selection method (selection with pac or bsd in vitro).
For the tgloII targeting vector, pBS/5tgoII-5E3DPacPB-3tgloII, recombination arms obtained by PCR were sequentially cloned into the pBluescript SK + plasmid pacPB expression cassette [19]. For the cgloII targeting vector, pBS/5cgoII-5E3DBsdPB-3cgloII, recombination arms obtained by PCR were sequentially cloned into the pBluescript SK + plasmid bsdPB expression cassette
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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P.berghei model of malaria
Technical design and realization: S. Mededovic and A. van Wigcheren