RMgmDB - Rodent Malaria genetically modified Parasites

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Summary

RMgm-1445
Malaria parasiteP. yoelii
Genotype
TaggedGene model (rodent): PY17X_1108700; Gene model (P.falciparum): PF3D7_0508000; Gene product: 6-cysteine protein (P38; Pf38)
Name tag: triple-HA
Phenotype Sporozoite;
Last modified: 6 May 2016, 20:18
  *RMgm-1445
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) : 27128092
MR4 number
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
The mutant parasite was generated by
Name PI/ResearcherSwearingen KE; Kappe SH; Sinnis P
Name Group/DepartmentJohns Hopkins Malaria Research Institute and Department of Molecular Microbiology and Immunology
Name InstituteJohns Hopkins Bloomberg School of Public Health, Johns Hopkins University
CityBaltimore
CountryUSA
Name of the mutant parasite
RMgm numberRMgm-1445
Principal nameHA(3x)-PY17X_1108700
Alternative name
Standardized name
Is the mutant parasite cloned after genetic modificationNo
Phenotype
Asexual blood stageNot tested
Gametocyte/GameteNot tested
Fertilization and ookineteNot tested
OocystNot tested
SporozoiteThe staining and localization of the 6-Cys protein p38 was internal to CSP and matched that of TRAP, a known micronemal protein that is secreted during motility and translocated to the sporozoite surface.
Liver stageNot tested
Additional remarks phenotype

Mutant/mutation
The mutant expresses a C-terminal 3xHA-tagged version of P38 (PY17X_1108700)

Protein (function)
The P38 protein is a member of a small family of proteins, the 6-cysteine (cys) family of (surface) proteins. The proteins are characterized by domains of roughly 120 amino acids in size that contain six positionally conserved cysteines (6-cys). Although some species of Plasmodium (may) contain unique members of the 6-cys family, ten members have been identified that are conserved both in structure as well as in genome organization throughout the genus. Some of the conserved 6-cys proteins are encoded by genes that form paralogous gene-pairs which are closely linked in the genome separated by less then 2 kb of intergenic region. Most members have a GPI anchor and are predicted membrane surface proteins whereas others appear to be secreted and most members are expressed in a discrete stage-specific manner in gametocytes, sporozoites or merozoites (see also 'Additional Information'). Evidence has been presented that P38 is transcribed in both asexual blood stages and in gametocytes.

Phenotype
In sporozoites the staining and localization of P38 was internal to CSP and matched that of TRAP, a known micronemal protein that is secreted during motility and translocated to the sporozoite surface.

Additional information

Other mutants
RMgm-354: a P. berghei mutant lacking expression of P38


  Tagged: Mutant parasite with a tagged gene
Details of the target gene
Gene Model of Rodent Parasite PY17X_1108700
Gene Model P. falciparum ortholog PF3D7_0508000
Gene product6-cysteine protein
Gene product: Alternative nameP38; Pf38
Details of the genetic modification
Name of the tagtriple-HA
Details of taggingC-terminal
Additional remarks: tagging
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 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 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