Summary

RMgm-4427
Malaria parasiteP. berghei
Genotype
DisruptedGene model (rodent): PBANKA_0828100; Gene model (P.falciparum): PF3D7_0927300; Gene product: fumarate hydratase (FH)
PhenotypeNo phenotype has been described
Last modified: 8 March 2018, 18:30
  *RMgm-4427
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) : 29449371
MR4 number
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
The mutant parasite was generated by
Name PI/ResearcherJayaraman V, Balaram H
Name Group/DepartmentJawaharlal Nehru Centre for Advanced Scientific Research
Name InstituteJawaharlal Nehru Centre for Advanced Scientific Research
City-
CountryIndia
Name of the mutant parasite
RMgm numberRMgm-4427
Principal name-
Alternative name
Standardized name
Is the mutant parasite cloned after genetic modificationNo
Phenotype
Asexual blood stageNot tested
Gametocyte/GameteNot tested
Fertilization and ookineteNot tested
OocystNot tested
SporozoiteNot tested
Liver stageNot tested
Additional remarks phenotype

Mutant/mutation
Selection of knock-out mutants was unsuccessful when transfected parasites were grown and selected in BALB/c mice. In C57Bl6 mice pyrimethamine-resistant populations were selected. PCR analyses provided evidence for disruption of the FH-gene. The mutants were not cloned and further analysed for phenotypes. These experiments may provide some evidence that FH is not essential for blood stage growth.

Protein (function)
Plasmodium has an iron-sulfur cluster–containing class I fumarate hydratase (FH) that catalyzes the interconversion of fumarate to malate, a well-known reaction in the tricarboxylic acid cycle.
Fumarate is generated in large quantities in the parasite as a byproduct of AMP synthesis and is converted to malate by FH and then used in the generation of the key metabolites oxaloacetate, aspartate, and pyruvate.

Phenotype
Selection of knock-out mutants was unsuccessful when transfected parasites were grown and selected in BALB/c mice. In C57Bl6 mice pyrimethamine-resistant populations were selected. PCR analyses provided evidence for disruption of the FH-gene. The mutants were not cloned and further analysed for phenotypes. These experiments may provide some evidence that FH is not essential for blood stage growth.

Additional information
Evidence is provided for mitochondrial localisation of FH

Other mutants


  Disrupted: Mutant parasite with a disrupted gene
Details of the target gene
Gene Model of Rodent Parasite PBANKA_0828100
Gene Model P. falciparum ortholog PF3D7_0927300
Gene productfumarate hydratase
Gene product: Alternative nameFH
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 usedYes
Name of PlasmoGEM construct/vector-
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 parasitehdhfr/yfcu
Promoter of the selectable markereef1a
Selection (positive) procedurepyrimethamine
Selection (negative) procedureNo
Additional remarks genetic modificationPlamoGem construct PbG01-2466a09 was used
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