Summary

RMgm-4065
Malaria parasiteP. berghei
Genotype
DisruptedGene model (rodent): PBANKA_1119200; Gene model (P.falciparum): PF3D7_0620000; Gene product: secreted ookinete protein 25, putative | ookinete surface-associated protein 8, putative (PSOP25, PSOP8)
Phenotype Gametocyte/Gamete; Fertilization and ookinete; Oocyst;
Last modified: 9 January 2017, 16:49
  *RMgm-4065
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) : 28057055
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/ResearcherZheng W, Cui L
Name Group/DepartmentDepartment of Entomology
Name InstituteThe Pennsylvania State University
CityUniversity Park, PA
CountryUSA
Name of the mutant parasite
RMgm numberRMgm-4065
Principal nameΔpsop25
Alternative name
Standardized name
Is the mutant parasite cloned after genetic modificationYes
Phenotype
Asexual blood stageNot different from wild type
Gametocyte/GameteNormal gametocyte numbers and sex ratio.
Mean male gamete exflagellation events were slightly but significantly reduced in the Δpsop25 line, as compared to the WT parasites.
Fertilization and ookineteSignificantly reduced ookinete production (60% in vitro). Mature ookinetes formed showed normal (light microscope) morphology.
OocystSignificantly reduced oocyst production (69%)
SporozoiteNot tested
Liver stageNot tested
Additional remarks phenotype

Mutant/mutation
The mutant lacks expression of PSOP25

Protein (function)
Psop25 encodes a 350 amino acid (aa) protein with a signal peptide, and the native protein is predicted to be 40 kDa. Psop25 transcript is highly expressed in ookinetes. The protein is expressed in ookinetes. The protein is called PSOP8 in another paper (see RMgm-1299).

Phenotype
Normal gametocyte numbers and sex ratio. Mean male gamete exflagellation events were slightly but significantly reduced in the Δpsop25 line, as compared to the WT parasites.
Significantly reduced ookinete production (60% in vitro). Mature ookinetes formed showed normal (light microscope) morphology.
Significantly reduced oocyst production (69%).

See also RMgm-1299 for an independent mutant lacking PSOP25/PSOP8. This mutant showed normal ookinete production but strong reduction of oocyst production (>20 fold).

Additional information
IFA showed the preferential expression of PSOP25 on the surface of zygotes, retorts and mature ookinetes.
Polyclonal and monoclonal antibodies against PSOP25 could effectively block the formation of ookinetes in vitro and transmission of the parasites to mosquitoes.

Other mutants
See PBANKA_1119200
 


  Disrupted: Mutant parasite with a disrupted gene
Details of the target gene
Gene Model of Rodent Parasite PBANKA_1119200
Gene Model P. falciparum ortholog PF3D7_0620000
Gene productsecreted ookinete protein 25, putative | ookinete surface-associated protein 8, putative
Gene product: Alternative namePSOP25, PSOP8
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/vector042760
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