Summary

RMgm-34
Malaria parasiteP. berghei
Genotype
DisruptedGene model (rodent): PBANKA_0209100; Gene model (P.falciparum): PF3D7_0104000; Gene product: thrombospondin-related sporozoite protein|sporozoite stage TSP1 domain protein (thrombospondin related sporozoite protein; TRSP; Sporozoite stage TSP1 domain protein, S21)
Phenotype Sporozoite; Liver stage;
Last modified: 12 February 2009, 13:18
  *RMgm-34
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) : 17418435
MR4 number
Parent parasite used to introduce the genetic modification
Rodent Malaria ParasiteP. berghei
Parent strain/lineP. berghei NK65
Name parent line/clone Not applicable
Other information parent line
The mutant parasite was generated by
Name PI/ResearcherM. Labaied, S.H.I. Kappe
Name Group/DepartmentDepartment of Pathobiology
Name InstituteUniversity of Washington
CitySeattle
CountryUSA
Name of the mutant parasite
RMgm numberRMgm-34
Principal namePbtrsp(−)
Alternative name
Standardized name
Is the mutant parasite cloned after genetic modificationYes
Phenotype
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
SporozoiteNormal numbers of salivary gland sporozoites are produced. Sporozoites show a gliding motility similar to wild type sporozoites. Invasion of hepatocytes is significantly reduced (see phenotype 'liver stage').
Liver stageIn vitro invasion of hepatocytes by the sporozoites is significantly reduced (35-38% reduction in number of completely intracellular sporozoites compared to wild type sporozoites). Invaded sporozoites develop normally inside HepG2 cells into mature liver schizonts.
In vivo a 40-80% reduction in liver infection is observed after infection of C57BL/6 mice by intraveneous inoculation of sporozoites.
Additional remarks phenotype

The mutant lacks expression of the thrombospondin related sporozoite protein (TRSP), a thrombospondin type 1 repeat-containing protein (TRS, TSP1). The phenotype analysis indicates a role of this protein in heptocyte entry.


  Disrupted: Mutant parasite with a disrupted gene
Details of the target gene
Gene Model of Rodent Parasite PBANKA_0209100
Gene Model P. falciparum ortholog PF3D7_0104000
Gene productthrombospondin-related sporozoite protein|sporozoite stage TSP1 domain protein
Gene product: Alternative namethrombospondin related sporozoite protein; TRSP; Sporozoite stage TSP1 domain protein, S21
Details of the genetic modification
Inducable system usedNo
Additional remarks inducable system
Type of plasmid/construct usedPlasmid 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 parasitetgdhfr
Promoter of the selectable markerpbdhfr
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 15'-TAATGGTACCGTTTGTGCACCTTAATCCC-3'
Additional information primer 1PbTRSPRep-5'UTR-For
Sequence Primer 25'-TAATAAGCTTCATATAAACTATGAACG-3'
Additional information primer 2PbTRSPRep-5'UTR-Rev
Sequence Primer 35'-TAATGAATTCGATAATAGTTTGGCTTTTCG-3'
Additional information primer 3PbTRSPRep-3'UTR-For
Sequence Primer 45'-TAATGGATCCGCAAAATATAGTCTACACCC-3'
Additional information primer 4PbTRSPRep-3UTR-Rev
Sequence Primer 5
Additional information primer 5
Sequence Primer 6
Additional information primer 6