RMgmDB - Rodent Malaria genetically modified Parasites

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Summary

RMgm-837
Malaria parasiteP. berghei
Genotype
DisruptedGene model (rodent): PBANKA_1113000; Gene model (P.falciparum): PF3D7_0513300; Gene product: purine nucleoside phosphorylase (PNP)
PhenotypeNo phenotype has been described
Last modified: 11 January 2018, 17:54
  *RMgm-837
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) : 23402751
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/ResearcherNiikura, M; Yuda, M; Kobayashi , F
Name Group/DepartmentDepartment of Infectious Diseases
Name InstituteKyorin University School of Medicine
CityTokyo
CountryJapan
Name of the mutant parasite
RMgm numberRMgm-837
Principal nameΔpbpnp
Alternative name
Standardized name
Is the mutant parasite cloned after genetic modificationYes
Phenotype
Asexual blood stageNot different from wild type
Gametocyte/GameteNot tested
Fertilization and ookineteNot tested
OocystNot tested
SporozoiteNot tested
Liver stageNot tested
Additional remarks phenotype

Mutant/mutation
The mutant lacks expression of purine nucleoside phosphorylase (PNP) and expresses GFP under the control of the P. berghei hsp70 promoter.

Protein (function)
Plasmodium species are unable to synthesize purine rings de novo; instead, they rely on purine nucleosides from the host. During the asexual phase of malaria parasites, adenosine is converted to inosine by adenosine deaminase (ADA) in the purine salvage pathway. Hypoxanthine is produced from inosine by purine nucleoside phosphorylase (PNP).

Phenotype
Normal growth and multiplication of asexual blood stages. Parasites induce experimental cerebral complications in C57BL/6 mice.

The normal growth of parasites lacking PNP suggests that these parasites are able to efficiently obtain hypoxanthine from other sources

Additional information
See also RMgm-838  for a P. yoelii YM mutant lacking expression of PNP. This mutant  shows attenuation of growth of blood stages and mice are able to resolve infections. In addition, this mutant did not produce oocysts.

Other mutants
RMgm-838: P. yoelii YM mutant lacking expression of PNP


  Disrupted: Mutant parasite with a disrupted gene
Details of the target gene
Gene Model of Rodent Parasite PBANKA_1113000
Gene Model P. falciparum ortholog PF3D7_0513300
Gene productpurine nucleoside phosphorylase
Gene product: Alternative namePNP
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 KpnI, NotI
Partial or complete disruption of the geneComplete
Additional remarks partial/complete disruption
Selectable marker used to select the mutant parasitehdhfr
Promoter of the selectable markereef1a
Selection (positive) procedurepyrimethamine
Selection (negative) procedureNo
Additional remarks genetic modificationΔpbpnp parasites were generated by double-crossover homologous recombination. In Δpbpnp parasites, a selection cassette containing gfp and an hDHFR–ts fusion gene was integrated into the target gene (pbpnp)
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 1AAAGGTACCTAAAATCCACGTAACCGCCA
Additional information primer 1PNP–5’F
Sequence Primer 2AAACTCGAGTCCAGGATCGCCAACAACTA
Additional information primer 2PNP–5’R
Sequence Primer 3AAAGGATCCATGAAGGGGATTTTGACAATC
Additional information primer 3PNP–3’F
Sequence Primer 4AAAGCGGCCGCGGATTTCATAAATGAGCTA
Additional information primer 4PNP–3’R
Sequence Primer 5
Additional information primer 5
Sequence Primer 6
Additional information primer 6