RMgmDB - Rodent Malaria genetically modified Parasites

Summary

RMgm-856
Malaria parasiteP. yoelii
Genotype
Transgene
 Transgene not Plasmodium: GFP
Promoter: Gene model: PBANKA_1133300; Gene model (P.falciparum): PF3D7_1357100; Gene product: elongation factor 1-alpha (eef1a)
3'UTR: Gene model: PBANKA_0711900; Gene product: heat shock protein 70 (HSP70)
Phenotype Asexual bloodstage;
Last modified: 4 April 2013, 10:53
  *RMgm-856
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) : 23535896
MR4 number
top of page
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
top of page
The mutant parasite was generated by
Name PI/ResearcherImai, T; Hisaeda H
Name Group/DepartmentDepartment of Parasitology
Name InstituteGraduate School of Medicine, Gunma University
CityGunma
CountryJapan
top of page
Name of the mutant parasite
RMgm numberRMgm-856
Principal namePyNL-GFP
Alternative name
Standardized name
Is the mutant parasite cloned after genetic modificationNo
top of page
Phenotype
Asexual blood stageGFP expression in blood stages
Gametocyte/GameteNot tested
Fertilization and ookineteNot tested
OocystNot tested
SporozoiteNot tested
Liver stageNot tested
Additional remarks phenotype

Mutant/mutation
The mutants expresses GFP under control of the constitutive eef1a promoter. The GFP-expression cassette is introduced on a Plasmodium artificial chromosome (PAC)

PyNL-GFP was generated using a PAC as described by Iwanaga et al (2010, Cell Host & Microbe 7, 245-255) . The following elements were inserted into the PAC: 1) elongation factor promoter; 2) green fluorescence protein (GFP); 3) Pb hsp70 3'-untranslated region; 4) P. berghei (Pb) pyrimethamine resistant gene dihydrofolate reductase-thymidyltransferase-ts (DHFR-ts) gene; 5) Pb DHFR-ts 3'-untranslated region.

In the paper no details are provided on (the generation of) the PAC such as centromeric (and telomeric regions; P. berghei or P. yoelii sequences?); circular or linear PAC. In addition no primer sequences (and gene IDs) are provided for amplification of the different regulatory sequences used.

The PAC used has been introduced as a circular PAC containing the P. yoelii chromosome 5 centromeric region.

  Transgene: Mutant parasite expressing a transgene
top of page
Type and details of transgene
Is the transgene Plasmodium derived Transgene: not Plasmodium
Transgene nameGFP
top of page
Details of the genetic modification
Inducable system usedNo
Additional remarks inducable system
Type of plasmid/constructPlasmodium Artificial Chromosome (PAC) circulair
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 parasitepbdhfr
Promoter of the selectable markereef1a
Selection (positive) procedurepyrimethamine
Selection (negative) procedureNo
Additional remarks genetic modificationPyNL-GFP was generated using a PAC as described by Iwanaga et al (2010, Cell Host & Microbe 7, 245-255) . The following elements were inserted into the PAC: 1) elongation factor promoter; 2) green fluorescence protein (GFP); 3) Pb hsp70 3'-untranslated region; 4) P. berghei (Pb) pyrimethamine resistant gene dihydrofolate reductase-thymidyltransferase-ts (DHFR-ts) gene; 5) Pb DHFR-ts 3'-untranslated region.

In the paper no details are provided on (the generation of) the PAC such as centromeric (and telomeric regions; P. berghei or P. yoelii sequences?); circular or linear PAC. In addition no primer sequences (and gene IDs) are provided for amplification of the different regulatory sequences used.

The PAC used has been introduced as a circular PAC containing the P. yoelii chromosome 5 centromeric region.
Additional remarks selection procedure
top of page
Other details transgene
top of page
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
top of page
3'-UTR
Gene Model of Parasite PBANKA_0711900
Gene productheat shock protein 70
Gene product: Alternative nameHSP70
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 / InsertionNot available
Gene Model of Parasite Not available
Gene productNot available
Gene product: Alternative name
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
top of page
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