SummaryRMgm-5229
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Successful modification | The 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) : 36126089 |
MR4 number | |
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Parent parasite used to introduce the genetic modification | |
Rodent Malaria Parasite | P. berghei |
Parent strain/line | P. berghei ANKA |
Name parent line/clone | unknown |
Other information parent line | |
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The mutant parasite was generated by | |
Name PI/Researcher | Gabelich JA, Ingmundson A |
Name Group/Department | Molecular Parasitology |
Name Institute | Humboldt University |
City | Berlin |
Country | Germany |
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Name of the mutant parasite | |
RMgm number | RMgm-5229 |
Principal name | PBANKA_1400700-mCherry |
Alternative name | |
Standardized name | |
Is the mutant parasite cloned after genetic modification | Yes |
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Phenotype | |
Asexual blood stage | PBANKA_1400700 localizes to punctate structures in the cytoplasm of infected red blood cells |
Gametocyte/Gamete | Not tested |
Fertilization and ookinete | Not tested |
Oocyst | Not tested |
Sporozoite | Not tested |
Liver stage | PBANKA_1400700-mCherry was detected throughout the liver stage, but it was not secreted from the parasite despite the presence of a signal peptide. |
Additional remarks phenotype | Mutant/mutation Protein (function) Additional information - IPIS2 and IPIS3 localizes to punctate structures in the cytoplasm of infected red blood cells In order to determine if the putative orthologs from P. vivax also targeted the membrane structures in the cytoplasm of Plasmodium-infected red blood cells, we generated P. berghei lines that expressed HA-tagged PvTRAg8, the syntenic P. vivax ortholog of IPIS2 (PVP01_0532600) or HA-tagged PvTRAg2, which shares similarity to IPIS3 (PVP01_0202200). These genes were integrated into the P. berghei genome such that they are expressed under the control of the IPIS2 or IPIS3 promoters, respectively. Both PvTRAg8-HA and PvTRAg2-HA are exported efficiently by P. berghei and localize to discrete structures in the cytoplasm of infected erythrocytes. Other mutants |
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Details of the target gene | |||||||||||||||||||||||||||
Gene Model of Rodent Parasite | PBANKA_1400700 | ||||||||||||||||||||||||||
Gene Model P. falciparum ortholog | Not available | ||||||||||||||||||||||||||
Gene product | Plasmodium exported protein, unknown function | ||||||||||||||||||||||||||
Gene product: Alternative name | |||||||||||||||||||||||||||
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Details of the genetic modification | |||||||||||||||||||||||||||
Name of the tag | mCherry | ||||||||||||||||||||||||||
Details of tagging | C-terminal | ||||||||||||||||||||||||||
Additional remarks: tagging | |||||||||||||||||||||||||||
Commercial source of tag-antibodies | |||||||||||||||||||||||||||
Type of plasmid/construct | (Linear) plasmid double cross-over | ||||||||||||||||||||||||||
PlasmoGEM (Sanger) construct/vector used | No | ||||||||||||||||||||||||||
Modified PlasmoGEM construct/vector used | No | ||||||||||||||||||||||||||
Plasmid/construct map | |||||||||||||||||||||||||||
Plasmid/construct sequence | |||||||||||||||||||||||||||
Restriction sites to linearize plasmid | |||||||||||||||||||||||||||
Selectable marker used to select the mutant parasite | tgdhfr | ||||||||||||||||||||||||||
Promoter of the selectable marker | pbdhfr | ||||||||||||||||||||||||||
Selection (positive) procedure | pyrimethamine | ||||||||||||||||||||||||||
Selection (negative) procedure | No | ||||||||||||||||||||||||||
Additional remarks genetic modification | For the construction of the plasmids used for endogenous C-terminal mCherry tagging, regions containing the IPIS2, IPIS3 or PBANKA_1400700 genes were amplified from P. berghei genomic DNA using the respective primers and integrated into the SacII and XbaI sites of the B3D+mCherry plasmid. The sequences of PvTRAg8 and PvTRAg2 were codon-optimized synthesized (GenScript) and cloned into the NotI and SpeI sites of b3D.DT^H.^D. The 5’ and 3’ untranslated regions of the IPIS2 and IPIS3 genes were cloned into the GOMO-GFP luciferase vector using the respective primers containing SacII and NotI sites and the XhoI and KpnI sites, respectively, in order to delete IPIS2 and IPIS3 via double homologous recombination. | ||||||||||||||||||||||||||
Additional remarks selection procedure | |||||||||||||||||||||||||||
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