RMgmDB - Rodent Malaria genetically modified Parasites

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Summary

RMgm-1252
Malaria parasiteP. berghei
Genotype
DisruptedGene model (rodent): PBANKA_0701000; Gene model (P.falciparum): PF3D7_0830500; Gene product: sporozoite and liver stage tryptophan-rich protein, putative | tryptophan-rich antigen (TryThrA; SLTRiP)
Transgene
Transgene not Plasmodium: GFP
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_0701000; Gene product: sporozoite and liver stage tryptophan-rich protein, putative (TryThrA; SLTRiP)
Phenotype Liver stage;
Last modified: 27 July 2015, 18:19
  *RMgm-1252
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) : 25960542
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/ResearcherJaijyan DK, Singh H, Singh AP
Name Group/DepartmentInfectious Diseases Laboratory
Name InstituteNational Institute of Immunology, Aruna Asaf Ali Marg
CityNew Delhi
CountryIndia
Name of the mutant parasite
RMgm numberRMgm-1252
Principal namePbĪ”SLTRiP::hDHFR&GFP
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 tested
OocystNot different from wild type
SporozoiteNot different from wild type
Liver stageReduction of the number of liver stages in vivo (21-fold fewer) and in vitro. One day delay in prepatent period in mice infected with mutant sporozoites. Evidence is presented for a reduced size of liver stages.
Additional remarks phenotype

Mutant/mutation
The mutant lacks expression of SLTRiP (and expresses GFP under the control of an unknown promoter)

Protein (function)
Sporozoite and liver stage tryptophan-rich protein, putative.
Different Plasmodium species express multiple tryptophan-rich proteins which are encoded by genes located near/in subtelomeric regions that often do not have a syntenic location between different species. The orthology between genes of different Plasmodium species is therefore less clear. See also PBANKA_052430 (tryptophan/threonine-rich antigen, putative) and RMgm-488, RMgm-729

Multiple sequence alignments shows that a segment of SLTRiP is conserved in Plasmodium species and contains a large number of position-constrained conserved tryptophan residues SLTRiP also contains a few cysteines (3 in P. berghei), but these are not conserved among other species.

In GeneDB (www.genedb.org) the PBANKA_070100 gene has been annotated as pseudogene (containing a  frameshift).

Phenotype
Normal blood stage development and production of oocysts and sporozoites. Reduction of the number of liver stages in vivo (21-fold fewer) and in vitro. One day delay in prepatent period in mice infected with mutant sporozoites. Evidence is presented for a reduced size of liver stages.

Additional information
Immunofluorescence evidence is presented for expression in sporozoites and liver stages and for export into the cytoplasm of the infected hepatocyte. No expression was detected in blood stages.

Other mutants


  Disrupted: Mutant parasite with a disrupted gene
Details of the target gene
Gene Model of Rodent Parasite PBANKA_0701000
Gene Model P. falciparum ortholog PF3D7_0830500
Gene productsporozoite and liver stage tryptophan-rich protein, putative | tryptophan-rich antigen
Gene product: Alternative nameTryThrA; SLTRiP
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 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 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
Promoter of the selectable markerpbdhfr
Selection (positive) procedurepyrimethamine
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_0701000
Gene productsporozoite and liver stage tryptophan-rich protein, putative
Gene product: Alternative nameTryThrA; SLTRiP
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