Summary

RMgm-237
Malaria parasiteP. berghei
Genotype
DisruptedGene model (rodent): PBANKA_1024600; Gene model (P.falciparum): PF3D7_1418100; Gene product: liver specific protein 1, putative (LISP1, liver specific protein 1)
Phenotype Liver stage;
Last modified: 23 May 2009, 23:21
  *RMgm-237
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) : 19438514
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/ResearcherT. Ishino, P. Baldacci
Name Group/DepartmentBiologie et Génétique du Paludisme
Name InstituteInstitut Pasteur
CityParis
CountryFrance
Name of the mutant parasite
RMgm numberRMgm-237
Principal nameNK65Lisp1Δ
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
SporozoiteNot different from wild type
Liver stageBy analysis of blood stage parasitemia after intravenous injection of sporozoites, the mutant sporozoites showed a 15-fold decrease in infectivity.
Additional remarks phenotype

Mutant/mutation
The mutant lacks expression of LISP1 (liver specific protein 1).

Protein (function)
Phenotype analyses of liver stage development of this and additional mutants lacking expression of LISP1 (RMgm-279, RMgm-280)  indicate that LISP plays a role in egress of merozoites from liver cells.

Phenotype
Phenotype analyses of liver stage development of additional mutants lacking expression of LISP1 (RMgm-279, RMgm-280)  indicate that LISP plays a role in egress of merozoites from liver cells, resulting from a defect in rupture of the membrane of the parasitophorous vacuole (PVM) and remain trapped inside hepatocytes.

Additional information
Using polyclonal antibody expression was detected in 48h liver schizonts in livers from rats and in infected HepG2 cells at 36, 48 and 65h post infection. No signal was detected in blood stages and sprorozoites. IFA analysis indicated a location of LISP in the parasitophorous vacuole membrane (PVM).
Four gene models exist: PB000708.00.0; PB000682.00.0; PB001247.00.0; PB000250.00.0.
Gen Bank accession no: AB231328.

Other mutants
RMgm-279: An independent mutant lacking expression of LISP1 and expressing GFP under the constitutive eef1a promoter
RMgm-280: An independent mutant lacking expression of LISP1 (ANKA strain)


  Disrupted: Mutant parasite with a disrupted gene
Details of the target gene
Gene Model of Rodent Parasite PBANKA_1024600
Gene Model P. falciparum ortholog PF3D7_1418100
Gene productliver specific protein 1, putative
Gene product: Alternative nameLISP1, liver specific protein 1
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 parasitehdhfr
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 15’- CGATGCGGGCCCGAGAATACAACTTACTAGGAAATGCAC-3’
Additional information primer 1ApaI - 630bp
Sequence Primer 25’- AGCTGGCGCGCCCTTGCATCTTCAGACATGTTATTTTCGA-3’
Additional information primer 2SmaI - 630bp
Sequence Primer 35’-
CCAGTGAGTGCGGCCGCGGCTAAAGCTCGATGTCGTATTCAAGAA-3’
Additional information primer 3NotI - 600bp
Sequence Primer 45’- AGCTGGCGCGCCCTGGGAATTTTTCAATTTCTTCCATTTG-3’
Additional information primer 4AscI - 600bp
Sequence Primer 5
Additional information primer 5
Sequence Primer 6
Additional information primer 6