RMgmDB - Rodent Malaria genetically modified Parasites


Malaria parasiteP. berghei
DisruptedGene model (rodent): PBANKA_1413600; Gene model (P.falciparum): PF3D7_1315100; Gene product: serine/threonine protein kinase PK9
PhenotypeNo phenotype has been described
Last modified: 26 May 2022, 17:39
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) : 35490588
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/ResearcherNarwal SK, Mishra S
Name Group/DepartmentDivision of Molecular Microbiology and Immunology
Name InstituteCSIR-Central Drug Research Institute
Name of the mutant parasite
RMgm numberRMgm-5206
Principal namePk9 KO
Alternative name
Standardized name
Is the mutant parasite cloned after genetic modificationYes
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

The mutant lacks expression of Pk9 and expresses GFP

Protein (function)
Previous studies have shown that functionally Pk9 can be designated as a serine-threonine kinase (STK) owing to its ability to phosphorylate E2 conjugation enzyme at serine 106 residue of Ubc13, although it does not cluster with any established eukaryotic protein kinase groups and hence is referred to as orphan kinase.

Normal, wild-type-like development throughout the complete life cycle

Additional information
 In this study also evidence is presented that that phosphorylation of the regulatory serine 106 of Ubc13 is essential for the completion of the parasite’s erythrocytic cycle. However,  phosphorylated form of Ubc13 was observed in Pk9 knockout parasites.

The process of ubiquitylation typically requires three enzymes: E1 (ubiquitin-activating enzyme, Uba), E2 (ubiquitin-conjugating enzyme, Ubc), and E3 (ubiquitin-protein ligase). Amongst Ubcs, Ubc13 specifically catalyzes the formation of K63-linked ubiquitin chains that are critical to signaling in inflammatory and DNA damage response pathways.

See also mutant RMgm-5208 with a mutated UNC13 (the phosphorylating serine 106 replaced with with non-phosphorylatable alanine).

Other mutants

  Disrupted: Mutant parasite with a disrupted gene
Details of the target gene
Gene Model of Rodent Parasite PBANKA_1413600
Gene Model P. falciparum ortholog PF3D7_1315100
Gene productserine/threonine protein kinase PK9
Gene product: Alternative name
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 markereef1a
Selection (positive) procedurepyrimethamine
Selection (negative) procedureNo
Additional remarks genetic modificationFor the deletion of Pk9, a replacement construct was generated as described previously (Mastan et al., 2017). For this, target sequences F8 and F9 were amplified using primers 1016/1017 and 1018/1019 specific for the 5′ or 3′ flanking regions respectively. The F8 (0.62 kb) and F9 (0.62 kb) were cloned in plasmid pBC-GFP-hDHFR at XhoI/ClaI and NotI/AscI restriction sites respectively. The targeting construct was digested with XhoI/AscI
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