RMgmDB - Rodent Malaria genetically modified Parasites


Malaria parasiteP. berghei
DisruptedGene model (rodent): PBANKA_0208300; Gene model (P.falciparum): PF3D7_0104800; Gene product: novel putative transporter 1, putative (Novel Putative Transporter, NPT1)
Phenotype Gametocyte/Gamete;
Last modified: 25 February 2017, 13:05
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) : 21752110
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/ResearcherRajendran E; van Dooren GG
Name Group/DepartmentResearch School of Biology
Name InstituteAustralian National University
Name of the mutant parasite
RMgm numberRMgm-4130
Principal nameΔPbnpt1
Alternative name
Standardized name
Is the mutant parasite cloned after genetic modificationYes
Asexual blood stageNot different from wild type
Gametocyte/GameteStrongly reduced formation of male gametes
Fertilization and ookineteNot tested
OocystNot tested
SporozoiteNot tested
Liver stageNot tested
Additional remarks phenotype

The mutant lacks expression of Novel Putative Transporter, NPT1. See also RMgm-626 for a more detailed analysis of a P. berghei mutant lacking NPT1.

Protein (function)
The P. falciparum homolog of P. berghei NPT1 has been reported to belong to an uncharacterized family of 5 novel putative transporters (NPTs) presenting some features of the Major Facilitating Superfamily (MFS) (Martin et al., 2005, Genome Biol.).  Plasmodium NPT1 proteins are predicted to contain 12 transmembrane domains. Transcription/expression of the npt1 gene has been shown for blood-, mosquito- and liver stages with increased transcript levels in liver stages.

See also RMgm-626 for a more detailed analysis of a P. berghei mutant lacking NPT1. Phenotype analyses of mutant RMgm-626 showed the following:  a strongly reduced gametocyte production. The few gametocytes present show an aberrant morphology (as shown by both ligh-microscopy and transmission electron microscopy analyses). These aberrant forms are unable to fertilize and to produce ookinetes.

Analysis of ΔPbnpt1 showed the following: Evidence is provided that blood stages of the mutant show reduced uptake of cationic amino acids (Arginine, Lysine). Evidence is presented that NPT1 is a cationic amino transporter in both T. gondii and Plasmodium.

Additional information
From the Abstract: 'Apicomplexans are obligate intracellular parasites that scavenge essential nutrients from their hosts via transporter proteins on their plasma membrane. The identities of the transporters that mediate amino acid uptake into apicomplexans are unknown. Here we demonstrate that members of an apicomplexan-specific protein family—the Novel Putative Transporters (NPTs)—play key roles in the uptake of cationic amino acids. We show that an NPT from Toxoplasma gondii (TgNPT1) is a selective arginine transporter that is essential for parasite survival and virulence. We also demonstrate that a homologue of TgNPT1 from the malaria parasite Plasmodium berghei (PbNPT1), shown previously to be essential for the sexual gametocyte stage of the parasite, is a cationic amino acid transporter. This reveals a role for cationic amino acid scavenging in gametocyte biology.'

Other mutants
see NPT1

  Disrupted: Mutant parasite with a disrupted gene
Details of the target gene
Gene Model of Rodent Parasite PBANKA_0208300
Gene Model P. falciparum ortholog PF3D7_0104800
Gene productnovel putative transporter 1, putative
Gene product: Alternative nameNovel Putative Transporter, NPT1
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 parasitehdhfr
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 1cgatgcgggcccctggctaattttgctattcatcattttac
Additional information primer 1Upstream targeting region forward (ApaI)
Sequence Primer 2cgatgccccgggatttcgaaataaatatattttatattctcaaatatg
Additional information primer 2Upstream targeting region reverse (XmaI)
Sequence Primer 3ccagtgagtgcggccgcgggcctgatttattcattaacctttttagc
Additional information primer 3Downstream targeting region forward (NotI)
Sequence Primer 4agctggcgcgccacagcgtgggtaaaacgaccgt
Additional information primer 4Downstream targeting region reverse (AscI)
Sequence Primer 5
Additional information primer 5
Sequence Primer 6
Additional information primer 6