RMgmDB - Rodent Malaria genetically modified Parasites

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Summary

RMgm-664
Malaria parasiteP. berghei
Genotype
TaggedGene model (rodent): PBANKA_0926700; Gene model (P.falciparum): PF3D7_1121600; Gene product: exported protein 1 | circumsporozoite-related antigen | parasitophorous vacuole membrane antigen QF 116 (CRA; exported protein1, Exp 1; U43539 hepatocyte erythrocyte protein 17 kDa (HEP17))
Name tag: mCherry
Phenotype Liver stage;
Last modified: 21 September 2016, 17:19
  *RMgm-664
Successful modificationThe parasite was generated by the genetic modification
The mutant contains the following genetic modification(s) Gene tagging
Reference (PubMed-PMID number) Reference 1 (PMID number) : 21909271
MR4 number
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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
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The mutant parasite was generated by
Name PI/ResearcherS. Graewe; V. Heussler
Name Group/DepartmentMalaria Lab I, Department of Molecular Parasitology
Name InstituteBernhard Nocht Institute for Tropical Medicine
CityHamburg
CountryGermany
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Name of the mutant parasite
RMgm numberRMgm-664
Principal nameP. berghei-Exp1-mCherry
Alternative name
Standardized name
Is the mutant parasite cloned after genetic modificationYes
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Phenotype
Asexual blood stageNot tested
Gametocyte/GameteNot tested
Fertilization and ookineteNot tested
OocystNot tested
SporozoiteNot tested
Liver stagemCherry signals detected in developing liver stages at the parasitophorous vacuole membrane (PVM)
Additional remarks phenotype

Mutant/mutation
A mutant expressing an additional copy of Exp1 (PBANKA_092670) which is (C-terminal) tagged with mCherry. The additional copy is under the control of the promoter of PBANKA_100300 (sequestrin, putative).
In the paper hardly any information is provided  regarding technical details of the genetic modification. It is therefore unknown whether the additional copy is introduced as an episome or is integrated into the genome.

Protein (function)
Exp1 (HEP17; circumsporozoite-related antigen) is a protein that localizes to the parasitophorous vacuole membrane (PVM) of blood stages and liver stages

Phenotype
mCherry signals detected in developing liver stages at the parasitophorous vacuole membrane (PVM).

Additional information
Hardly any information is provided in the paper regarding technical details of the genetic modification.

In the paper it is mentioned that constitutive expression of mCherry-tagged Exp1 led to a developmental arrest of the parasite (in the paper it is not mentioned at which stage, blood- or liverstage, this developmental arrest occurs and which promoter had been used for constitutive expression).

In the paper Kaiser et al. (2016). Mol Microbiol (PMID: 27566438) a similar mutant has been described that expresses  an additional copy of Exp1 (PBANKA_0926700) which is (C-terminal) tagged with GFP. This mutant shows normal development of liver stages.

Other mutants

  Tagged: Mutant parasite with a tagged gene
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Details of the target gene
Gene Model of Rodent Parasite PBANKA_0926700
Gene Model P. falciparum ortholog PF3D7_1121600
Gene productexported protein 1 | circumsporozoite-related antigen | parasitophorous vacuole membrane antigen QF 116
Gene product: Alternative nameCRA; exported protein1, Exp 1; U43539 hepatocyte erythrocyte protein 17 kDa (HEP17)
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Details of the genetic modification
Name of the tagmCherry
Details of taggingC-terminal
Additional remarks: taggingHardly any information is provided in the paper regarding technical details of the genetic modification.
Commercial source of tag-antibodies
Type of plasmid/constructNot known
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 parasiteNot known
Promoter of the selectable markerNot known
Selection (positive) procedurepyrimethamine
Selection (negative) procedureNo
Additional remarks genetic modificationHardly any information is provided in the paper regarding technical details of the 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
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Conceptual design of the database: Dr. C.J. Janse (Leiden Malaria Research Group, Leiden, The Netherlands).
Technical design and realization: S. Mededovic and A. van Wigcheren