RMgmDB - Rodent Malaria genetically modified Parasites

Summary

RMgm-791
Malaria parasiteP. berghei
Genotype
DisruptedGene model (rodent): PBANKA_1430800; Gene model (P.falciparum): PF3D7_1215000; Gene product: thioredoxin peroxidase 2 (Trx-Px2; TPx2; TPx-2)
PhenotypeNo phenotype has been described
Last modified: 14 November 2012, 21:02
  *RMgm-791
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) : 23146411
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/ResearcherH. Masuda-Suganuma; S.I. Kawazu
Name Group/DepartmentNational Research Center for Protozoan Diseases
Name InstituteObihiro University of Agriculture and Veterinary Medicine
CityInada, Obihiro
CountryJapan
Name of the mutant parasite
RMgm numberRMgm-791
Principal nameTPx-2 KO
Alternative name
Standardized name
Is the mutant parasite cloned after genetic modificationYes
Phenotype
Asexual blood stageNot different from wild type
Gametocyte/GameteNot different from wild type
Fertilization and ookineteNot tested
OocystNot different from wild type
SporozoiteNot tested
Liver stageNot tested
Additional remarks phenotype

Mutant/mutation
The mutant lacks expression of thioredoxin peroxidase 2 (TPx-2).

Protein (function)
Malaria parasites possess six peroxidases localized in the cytoplasm, mitochondrion, apicoplast and nucleus: a 1-Cys Prx, two typical 2-Cys Prxs, a 1-Cys antioxidant protein (AOP), a Prx family with unusual biochemical characteristics and a glutathione (GSH) peroxidase-like thioredoxin peroxidase (TPxGl). Since malaria parasites do not possess catalase and genuine GSH peroxidase in their genome, it is considered that GSH itself is the major redox buffer for transient H2O2 exposure and the basal peroxide flux in the cell is dealt with by the Trx system.
 2-Cys Prxs have been found to act as a terminal peroxidase that reduces hydrogen peroxide and organic hydroperoxides with the use of electrons donated by the thioredoxin (Trx) system. Plasmodium contains 1-Cys Prx and two typical 2-Cys Prxs; TPx-1 (cytosolic) and TPx-2 (mitochondrial); see also 'Additional information').

Phenotype
Phenotype analyses indicate that TPx-2 is not essential for blood stage development and development of (gametocytes, ookinetes and) oocysts.

Additional information
In most eukaryotic organisms, redox-active enzymes, such as catalase, superoxide dismutase, and peroxidases as well as an enzymatic cascade that generates reduced electron donors, i.e. glutathione (GSH) and thioredoxin (Trx), sustain the cellular redox homeostasis. This redox network is split into two major arms, the GSH and the Trx system, that serve complementary functions in antioxidant defence and DNA synthesis. The malarial parasite Plasmodium lacks two central antioxidant enzymes: (i) catalase that typically detoxifies hydrogen peroxide and (ii) a classical glutathione peroxidase, a selenoenyzme that reduces lipid hydroperoxides to their alcohols. This apparent deficiency  underscores the possible central importance of the thioredoxin system in the parasite. 

See also mutant RMgm-206 which lacks expression of the 2-Cys peroxiredoxin, thioredoxin peroxidase 1 (TPx-1)

Other mutants
RMgm-206: A mutant lacking expression of thioredoxin peroxidase 1 (TPx-1)
RMgm-586: A mutant lacking expression of thioredoxin reductase (TrxR)


  Disrupted: Mutant parasite with a disrupted gene
Details of the target gene
Gene Model of Rodent Parasite PBANKA_1430800
Gene Model P. falciparum ortholog PF3D7_1215000
Gene productthioredoxin peroxidase 2
Gene product: Alternative nameTrx-Px2; TPx2; TPx-2
Details of the genetic modification
Inducable system usedNo
Additional remarks inducable system
Type of plasmid/construct usedPlasmid 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 parasitepbdhfr
Promoter of the selectable markerpbdhfr
Selection (positive) procedurepyrimethamine
Selection (negative) procedureNo
Additional remarks genetic modification
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 15′–GGG CCC CTA TCT GAG TAA TAT CTC ATA TCT CC-3′
Additional information primer 1
Sequence Primer 25′-GAG CTC CAT GAG AAA ACG ATC TCA CTG ATC-3′
Additional information primer 2
Sequence Primer 35′-GAA TTC GTT TAG ACC CAA TAA TGA GGC-3′
Additional information primer 3
Sequence Primer 45′-CTG CAG GTT TTT CAC ACA CAG GAG TTA C-3′
Additional information primer 4
Sequence Primer 5
Additional information primer 5
Sequence Primer 6
Additional information primer 6