Summary

RMgm-4830
Malaria parasiteP. berghei
Genotype
DisruptedGene model (rodent): PBANKA_1115200; Gene model (P.falciparum): PF3D7_0515600; Gene product: gamete egress protein GEP, putative (GEP)
Transgene
Transgene not Plasmodium: GFP
Promoter: Gene model: PBANKA_0711900; Gene model (P.falciparum): PF3D7_0818900; Gene product: heat shock protein 70 (GEP)
3'UTR: Gene model: Not available; Gene product: Not available
Replacement locus: Gene model: PBANKA_1115200; Gene product: gamete egress protein GEP, putative (GEP)
Phenotype Gametocyte/Gamete;
Last modified: 20 June 2020, 22:20
  *RMgm-4830
Successful modificationThe parasite was generated by the genetic modification
The mutant contains the following genetic modification(s) Gene disruption, Introduction of a transgene
Reference (PubMed-PMID number) Reference 1 (PMID number) : 32352241
MR4 number
Parent parasite used to introduce the genetic modification
Rodent Malaria ParasiteP. berghei
Parent strain/lineP. berghei ANKA
Name parent line/clone 8417HP
Other information parent line
The mutant parasite was generated by
Name PI/ResearcherAndreadaki M, Curra C
Name Group/DepartmentFORTH, Institute of Molecular Biology and Biotechnology
Name InstituteFORTH, Institute of Molecular Biology and Biotechnology
CityHeraklion
CountryGreece
Name of the mutant parasite
RMgm numberRMgm-4830
Principal namegep(‐) cl1, cl2
Alternative name
Standardized name
Is the mutant parasite cloned after genetic modificationYes
Phenotype
Asexual blood stageNot different from wild type
Gametocyte/GameteNormal production of male and female gametocytes. No exflagellation centers. Male and females are strongly impaired in gametocyte egress from the red blood cell (50% of the gep(‐) males had vesiculated PVM and in only 5.5% the RBCM ruptured. In females, only 4% of the gep(‐) females were fully released, while 38% had a ruptured PVM).
Fertilization and ookineteNot tested
OocystNot tested
SporozoiteNot tested
Liver stageNot tested
Additional remarks phenotype

Mutant/mutation
The mutant lacks expression of GEP and expresses GFP under control of the constitutive hsp70 promoter

Protein (function)
Gamete Egress Protein is a protein of 1,138 amino acids encoded by a 4,426 bp gene on chromosome 11 having 10 exons residing in a syntenic region

Phenotype
Normal production of male and female gametocytes. No exflagellation centers. Male and females are strongly impaired in gametocyte egress from the red blood cell (50% of the gep(‐) males had vesiculated PVM and in only 5.5% the RBCM ruptured. In females, only 4% of the gep(‐) females were fully released, while 38% had a ruptured PVM).

Additional information
Evidence is presented that:
- GEP is expressed in male and female gametocytes and partly localizes to osmiophillic bodies (OBs)
- Gep(‐) gametocytes are delayed in OB release upon activation of gametogenesis
- Gep(‐) male gametes form normal axonemes, but flagella are not motile
- Genetic crosses reveal a functional defect in both genders of gep(‐)

Other mutants


  Disrupted: Mutant parasite with a disrupted gene
Details of the target gene
Gene Model of Rodent Parasite PBANKA_1115200
Gene Model P. falciparum ortholog PF3D7_0515600
Gene productgamete egress protein GEP, putative
Gene product: Alternative nameGEP
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/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

  Transgene: Mutant parasite expressing a transgene
Type and details of transgene
Is the transgene Plasmodium derived Transgene: not Plasmodium
Transgene nameGFP
Details of the genetic modification
Inducable system usedNo
Additional remarks inducable system
Type of plasmid/construct(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
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
Other details transgene
Promoter
Gene Model of Parasite PBANKA_0711900
Gene Model P. falciparum ortholog PF3D7_0818900
Gene productheat shock protein 70
Gene product: Alternative nameGEP
Primer information details of the primers used for amplification of the promoter sequence  Click to view information
Primer information details of the primers used for amplification of the promoter sequence  Click to hide information
Sequence Primer 1
Additional information primer 1
Sequence Primer 2
Additional information primer 2
3'-UTR
Gene Model of Parasite Not available
Gene productNot available
Gene product: Alternative name
Primer information details of the primers used for amplification the 3'-UTR sequences  Click to view information
Primer information details of the primers used for amplification the 3'-UTR sequences  Click to hide information
Sequence Primer 1
Additional information primer 1
Sequence Primer 2
Additional information primer 2
Insertion/Replacement locus
Replacement / InsertionReplacement locus
Gene Model of Parasite PBANKA_1115200
Gene productgamete egress protein GEP, putative
Gene product: Alternative nameGEP
Primer information details of the primers used for amplification of the target sequences  Click to view information
Primer information details of the 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