RMgmDB - Rodent Malaria genetically modified Parasites

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Summary

RMgm-1512
Malaria parasiteP. yoelii
Genotype
TaggedGene model (rodent): PY17X_1457400; Gene model (P.falciparum): PF3D7_1241500; Gene product: conserved Plasmodium protein, unknown function
Name tag: c-myc
Phenotype Liver stage;
Last modified: 9 August 2016, 18:35
  *RMgm-1512
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) : 27434123
MR4 number
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Parent parasite used to introduce the genetic modification
Rodent Malaria ParasiteP. yoelii
Parent strain/lineP. y. yoelii 17XNL
Name parent line/clone Not applicable
Other information parent line
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The mutant parasite was generated by
Name PI/ResearcherSpeake C, Kappe SH, Krzych U, Duffy PE
Name Group/DepartmentLaboratory of Malaria Immunology and Vaccinology
Name InstituteNational Institute of Allergy and Infectious Diseases, National Institutes of Health
CityBethesda, Maryland
CountryUSA
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Name of the mutant parasite
RMgm numberRMgm-1512
Principal namePyPF3D7_1241500::myc
Alternative name
Standardized name
Is the mutant parasite cloned after genetic modificationNo
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Phenotype
Asexual blood stageNot tested
Gametocyte/GameteNot tested
Fertilization and ookineteNot tested
OocystNot tested
SporozoiteNot tested
Liver stageThe tagged protein was detected in 24h liver stages and localized to the parasite peri-nuclear area but was not exported.
Additional remarks phenotype

Mutant/mutation
The mutant expresses a cmyc-tagged (C-terminal) version of PY17X_1457400.

The gene was identified in a transcriptome-based approach to identify novel pre-erythrocytic vaccine antigens. A tiling microarray was used to identify P. falciparum genes transcribed at higher levels during liver stage versus sporozoite or blood stages of development.

Expression of the gene is only analysed in liver stages.
The tagged protein was detected in 24h liver stages and localized to the parasite peri-nuclear area but was not exported.

  Tagged: Mutant parasite with a tagged gene
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Details of the target gene
Gene Model of Rodent Parasite PY17X_1457400
Gene Model P. falciparum ortholog PF3D7_1241500
Gene productconserved Plasmodium protein, unknown function
Gene product: Alternative name
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Details of the genetic modification
Name of the tagc-myc
Details of taggingC-terminal
Additional remarks: taggingquadruple myc tag
Commercial source of tag-antibodies
Type of plasmid/construct(Linear) plasmid single 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 parasitetgdhfr
Promoter of the selectable markerpbdhfr
Selection (positive) procedurepyrimethamine
Selection (negative) procedureNo
Additional remarks genetic modificationThe b3D-myc vector for epitope tagging genes of interest was used to generate the Py
myc-tagged transgenic lines of PyPF3D7_0506200, PyPF3D7_1241500, and PyPF3D7_07
30200. Genes without their stop codons, together with approximately 1.5 kb sequence
upstream from the start codon, were amplified by PCR from Py 17XNL genomic DNA and
inserted upstream of the quadruple myc tag in b3D-myc.
The plasmids were subsequently linearized with Psr I and integrated into the Py 17XNL
genome using standard procedures. This integration strategy created two copies of the
target gene, both under the control of the endogenous promoter: the original gene and the
myc epitope-tagged gene.
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: StudioDrie StudioDrie; S. Mededovic and A. van Wigcheren