RMgmDB - Rodent Malaria genetically modified Parasites

Back to search results

Summary

RMgm-1170
Malaria parasiteP. berghei
Genotype
DisruptedGene model (rodent): PBANKA_1460700; Gene model (P.falciparum): PF3D7_1247800; Gene product: dipeptidyl aminopeptidase 2 (DPAP2)
PhenotypeNo phenotype has been described
Last modified: 17 February 2015, 19:16
  *RMgm-1170
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) : 23836185
MR4 number
top of page
Parent parasite used to introduce the genetic modification
Rodent Malaria ParasiteP. berghei
Parent strain/lineP. berghei ANKA
Name parent line/clone P. berghei ANKA 2.34
Other information parent lineP. berghei ANKA 2.34 is a cloned, gametocyte producer line of the ANKA strain (PubMed: PMID: 15137943).
top of page
The mutant parasite was generated by
Name PI/ResearcherTanaka TQ; Williamson KC
Name Group/DepartmentLaboratory of Malaria and Vector Research
Name InstituteNational Institute of Allergy and Infectious Diseases, National Institutes of Health
CityRockville
CountryUSA
top of page
Name of the mutant parasite
RMgm numberRMgm-1170
Principal namePbdpap2Δ
Alternative name
Standardized name
Is the mutant parasite cloned after genetic modificationYes
top of page
Phenotype
Asexual blood stageNot different from wild type
Gametocyte/GameteNot different from wild type
Fertilization and ookineteNot different from wild type
OocystNot different from wild type
SporozoiteNot different from wild type
Liver stageNot different from wild type
Additional remarks phenotype

Mutant/mutation
The mutant lacks expression of DPAP2

Protein (function)
The P. falciparum and P. berghei genomes each contain three dipeptidyl aminopeptidase (dpap) homologs. P. falciparum DPAP1 and DPAP3 are critical for asexual growth. The P. falciparum dpap2 gene could be deleted. P. falciparum parasites lacking expression of (the gametocyte-specific?) DPAP2 produced normal gametocytes.

In P. berghei blood stages, dpap2 transcription is highly upregulated in gametocytes compared to asexual blood stages.

DPAP3 of P. falciparum has been implicated in egress of merozoites from the host erythrocyte.

Evidence has been presented that the P. falciparum enzyme dipeptidyl aminopeptidase 1 (DPAP1; an ortholog of the lysosomal exopeptidase cathepsin C) was located in the food/digestive vacuole and to possesses hydrolytic activity against dipeptide substrates, suggesting that DPAP1 is involved in the generation of dipeptides from hemoglobin-derived oligopeptides. The dpap1 gene could not be deleted from the P. falciparum genome, suggesting an essential role during blood stage growth/multiplication.

In P. berghei all three dpap genes can be deleted from the genome (see below).

Phenotype
Phenotype analyses throughout the complete life cycle indicate that the mutant have growth and multiplication features that are similar to wild type parasites.

Additional information
The P. falciparum dpap2 gene could be deleted. P. falciparum parasites lacking expression of (the gametocyte-specific?) DPAP2 produced normal gametocytes.

Other mutants
RMgm-810: A mutant lacking expression of DPAP1
RMgm-812: A mutant lacking expression of DPAP3
RMgm-811: An independent mutant lacking expression of DPAP2
 

  Disrupted: Mutant parasite with a disrupted gene
top of page
Details of the target gene
Gene Model of Rodent Parasite PBANKA_1460700
Gene Model P. falciparum ortholog PF3D7_1247800
Gene productdipeptidyl aminopeptidase 2
Gene product: Alternative nameDPAP2
top of page
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 parasitetgdhfr
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’ TAAT GGGCCC TTTTAAGTTATACTATATTTTCCCT 3’
Additional information primer 1Pbdpap2.5’.Apa.F
Sequence Primer 25’ TAAT AAGCTT AATACGACGAGAGCTGGAATCATT 3’
Additional information primer 2Pbdpap2.5’.HindIII.R
Sequence Primer 35’ TCTAGA CACAGTACAATCAGGCATGTGATG 3’
Additional information primer 3Pbdpap2.3’.Xba.F
Sequence Primer 45’ CCGCGG CAAAATAACATCTATGAAATATGTATGAC 3’
Additional information primer 4Pbdpap2.3’.SacII.R
Sequence Primer 5
Additional information primer 5
Sequence Primer 6
Additional information primer 6
top of page
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