Summary

RMgm-4056
Malaria parasiteP. berghei
Genotype
DisruptedGene model (rodent): PBANKA_0512800; Gene model (P.falciparum): PF3D7_1028700; Gene product: merozoite TRAP-like protein (MTRAP)
Transgene
Transgene not Plasmodium: mCherrry
Promoter: Gene model: Not available; Gene model (P.falciparum): Not available; Gene product: Not available
3'UTR: Gene model: Not available; Gene product: Not available
Replacement locus: Gene model: PBANKA_0512800; Gene product: merozoite TRAP-like protein (MTRAP)
Phenotype Gametocyte/Gamete; Fertilization and ookinete; Oocyst;
Last modified: 15 November 2016, 17:29
  *RMgm-4056
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) : 27832590
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/ResearcherBargieri DY; Menard, R
Name Group/DepartmentMalaria Biology and Genetics Unit
Name InstitutePasteur Institute
CityParis
CountryFrance
Name of the mutant parasite
RMgm numberRMgm-4056
Principal namePbMTRAP(ko)
Alternative name
Standardized name
Is the mutant parasite cloned after genetic modificationYes
Phenotype
Asexual blood stageNot different from wild type
Gametocyte/GameteNormal numbers of gametocytes are produced. Both male and female gametes cannot escape from the host red blood cell after activation of gametocytes. Evidence is presented that the parasitophorous membrane is not disrupted in the absence of MTRAP
Fertilization and ookineteNo fertilisation and ookinetes. Normal numbers of gametocytes are produced. Both male and female gametes can escape from the hosr red blood cell after activation of gametocytes. Evidence is presented that the parasitophorous membrane is not disrupted in the absence of MTRAP
OocystNo oocyst production
SporozoiteNot tested
Liver stageNot tested
Additional remarks phenotype

Mutant/mutation
The mutant lacks expression of MTRAP and expresses mCherry (likely under control of a constitutive promoter but no details are provided on the promoter used)

Protein (function)
The thrombospondin-related anonymous protein (TRAP) family of proteins are type I transmembrane proteins that share a functionally conserved cytoplasmic tail and an ectodomain exposing various ligand-binding modules including a thrombospondin type I repeat (TSR). In Plasmodium, the sporozoite stage expresses three members of the family—TRAP; TRAP-related protein (TREP), also called S6; and TRAP-ike protein (TLP)—which all play a role in sporozoite gliding on substrates and within tissues. The ookinete stage expresses a single member, called circumsporozoite protein and thrombospondin-related anonymous protein-related protein (CTRP), which is essential for ookinete gliding motility.
Merozoite TRAP (MTRAP) is a TRAP family member that was reported as expressed in the merozoite. The mtrap gene is conserved and syntenic among Plasmodium species.

Phenotype
Normal numbers of gametocytes are produced. Both male and female gametes cannot escape from the host red blood cell after activation of gametocytes. Evidence is presented that the parasitophorous membrane is not disrupted in the absence of MTRAP.
No fertilisation and ookinete and oocyst production.

Additional information

Other mutants
See link


  Disrupted: Mutant parasite with a disrupted gene
Details of the target gene
Gene Model of Rodent Parasite PBANKA_0512800
Gene Model P. falciparum ortholog PF3D7_1028700
Gene productmerozoite TRAP-like protein
Gene product: Alternative nameMTRAP
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 markerunknown
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 namemCherrry
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
Promoter of the selectable markerunknown
Selection (positive) procedureNo
Selection (negative) procedureNo
Additional remarks genetic modification
Additional remarks selection procedure
Other details transgene
Promoter
Gene Model of Parasite Not available
Gene Model P. falciparum ortholog Not available
Gene productNot available
Gene product: Alternative name
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_0512800
Gene productmerozoite TRAP-like protein
Gene product: Alternative nameMTRAP
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