RMgmDB - Rodent Malaria genetically modified Parasites

Summary

RMgm-4640
Malaria parasiteP. berghei
Genotype
DisruptedGene model (rodent): PBANKA_1131900; Gene model (P.falciparum): PF3D7_1355500; Gene product: serine/threonine protein phosphatase 5 (PP5)
Phenotype Gametocyte/Gamete; Fertilization and ookinete; Oocyst; Sporozoite;
Last modified: 21 June 2019, 17:25
  *RMgm-4640
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) : 31202684
MR4 number
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).
The mutant parasite was generated by
Name PI/ResearcherZhu X, Cui L
Name Group/DepartmentDepartment of Immunology, College of Basic Medical Science
Name InstituteChina Medical University
CityShenyang
CountryChina
Name of the mutant parasite
RMgm numberRMgm-4640
Principal nameΔpbpp5
Alternative name
Standardized name
Is the mutant parasite cloned after genetic modificationYes
Phenotype
Asexual blood stageNot different from wild type
Gametocyte/GameteNormal (wild type) numbers of male and female gametocytes produced. Strong reduction in male gamete formation (exflagellation): 13.8 – 25% exflagellating Δpbpp5 male gametocytes compared with 88.9% in wild type parasites.
Fertilization and ookineteNo ookinete formation in vitro
OocystNo ookinete formation in vitro. Strong reducuction of oocyst formation (70-100%).
SporozoiteNo ookinete formation in vitro. Strong reducuction of oocyst formation (70-100%). No sporozoites detected in salivary glands
Liver stageNot tested
Additional remarks phenotype

Mutant/mutation
The mutant lacks expression of PP5

Protein (function)
Malaria parasites have homologues of all major human phosphoprotein phosphatase (PPP) subfamilies PP1 – PP7. Of the PPP subfamilies, PP5 differs from other phosphatases in possessing three N-terminal tetratricopeptide repeat (TPR) domains, which are auto-inhibited and responsible for protein-protein interactions. Being expressed in virtually all mammalian systems, PP5 interacts with a number of proteins through its TPR domains to function as a modulator in multiple signaling pathways.

Phenotype
Normal (wild type) numbers of male and female gametocytes produced. Strong reduction in male gamete formation (exflagellation): 13.8 – 25% exflagellating Δpbpp5 male gametocytes compared with 88.9% in wild type parasites.
No ookinete formation in vitro. Strong reducuction of oocyst formation (70-100%).
No sporozoites detected in salivary glands.
Crossing experiments showed that female Δpbpp5 gametocytes/gametes are fertile whereas Δpbpp5 male gametocyte/gamete fertility is strongly reduced.

Additional information
See RMgm-4641 for a mutant expressing C-terminal GFP-tagged version PP5: the PbPP5 protein was expressed in both blood stages and ookinetes with more prominent expression during sexual development. PbPP5 was localized in the cytoplasm of the parasite and highly concentrated beneath the parasite plasma membrane in free merozoites and ookinetes.

Other mutants


  Disrupted: Mutant parasite with a disrupted gene
Details of the target gene
Gene Model of Rodent Parasite PBANKA_1131900
Gene Model P. falciparum ortholog PF3D7_1355500
Gene productserine/threonine protein phosphatase 5
Gene product: Alternative namePP5
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 usedYes
Name of PlasmoGEM construct/vector301794
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/yfcu
Promoter of the selectable markereef1a
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 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