RMgmDB - Rodent Malaria genetically modified Parasites

Summary

RMgm-5221
Malaria parasiteP. yoelii
Genotype
Transgene
Transgene not Plasmodium: RFP (RedStar)
Promoter: Gene model: PBANKA_1133300; Gene model (P.falciparum): PF3D7_1357100; Gene product: elongation factor 1-alpha (eef1a)
3'UTR: Gene model: PBANKA_0719300; Gene product: bifunctional dihydrofolate reductase-thymidylate synthase, putative (dhfr/ts)
Replacement locus: Gene model: Not available; Gene product: Not available (small subunit ribosomal rna gene (c/d-type unit))
Phenotype Sporozoite; Liver stage;
Last modified: 15 July 2022, 16:12
  *RMgm-5221
Successful modificationThe parasite was generated by the genetic modification
The mutant contains the following genetic modification(s) Introduction of a transgene
Reference (PubMed-PMID number) Reference 1 (PMID number) : 27901110
MR4 number
Parent parasite used to introduce the genetic modification
Rodent Malaria ParasiteP. yoelii
Parent strain/lineP. y. yoelii 265BY
Name parent line/clone P. y. yoelii 265BY
Other information parent line265BY nonlethal strain (Py265)
The mutant parasite was generated by
Name PI/ResearcherZhao C, Xu W
Name Group/DepartmentDepartment of Pathogenic Biology
Name InstituteThird Military Medical University
CityChongqing
CountryChina
Name of the mutant parasite
RMgm numberRMgm-5221
Principal nameP. y. yoelii 265BY-RFP
Alternative name
Standardized name
Is the mutant parasite cloned after genetic modificationYes
Phenotype
Asexual blood stageNot tested
Gametocyte/GameteNot tested
Fertilization and ookineteNot tested
OocystNot tested
SporozoiteRFP expression in sporozoites
Liver stageRFP expression in liver stages
Additional remarks phenotype

Mutant/mutation
The mutant expresses RFP (redstar) under control of the constitutive eef1a promoter. The RFP expression cassette has been introduced by single cross-over integration into the small subunit ribosomal rna gene (c/d-type unit)

Protein (function)
Red fluorescent protein (RFP; RedStar)

Phenotype
RFP expression in sporozoites and in liver stages

Additional information

Other mutants


  Transgene: Mutant parasite expressing a transgene
Type and details of transgene
Is the transgene Plasmodium derived Transgene: not Plasmodium
Transgene nameRFP (RedStar)
Details of the genetic modification
Inducable system usedNo
Additional remarks inducable system
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 modificationP. y. yoelii 265BY-RFP was constructed by inserting RFP into the ssurRNA gene of P. y. yoelii 265BY.
In brief, PL1102 (a gift from MR4) was electroporated into cultured shizonts of P. y. yoelii 265BY and injected into a mouse.
One day later, the mice were treated with pyrimethamine in their drinking water for six days and parasites were collected and cloned.
Additional remarks selection procedure
Other details transgene
Promoter
Gene Model of Parasite PBANKA_1133300
Gene Model P. falciparum ortholog PF3D7_1357100
Gene productelongation factor 1-alpha
Gene product: Alternative nameeef1a
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 PBANKA_0719300
Gene productbifunctional dihydrofolate reductase-thymidylate synthase, putative
Gene product: Alternative namedhfr/ts
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 Not available
Gene productNot available
Gene product: Alternative namesmall subunit ribosomal rna gene (c/d-type unit)
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