SummaryRMgm-5265
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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) : 36625655 |
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 | Not applicable |
Other information parent line | |
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The mutant parasite was generated by | |
Name PI/Researcher | Wichers-Misterek JS, Gilberger TW |
Name Group/Department | Centre for Structural Systems Biology |
Name Institute | Centre for Structural Systems Biology |
City | Hamburg |
Country | Germany |
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Name of the mutant parasite | |
RMgm number | RMgm-5265 |
Principal name | PbSPM3-GFP |
Alternative name | |
Standardized name | |
Is the mutant parasite cloned after genetic modification | Yes |
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Phenotype | |
Asexual blood stage | Not different from wild type |
Gametocyte/Gamete | Not different from wild type |
Fertilization and ookinete | Not different from wild type |
Oocyst | PbSPM3-GFP expression in oocysts and sporozoites. In sporozoites, the fluorescent signal extended from the apical end towards the rear around the nucleus. |
Sporozoite | PbSPM3-GFP expression in oocysts and sporozoites. In sporozoites, the fluorescent signal extended from the apical end towards the rear around the nucleus. |
Liver stage | Not different from wild type |
Additional remarks phenotype | Mutant/mutation Protein (function) Phenotype For P. falciparum SPM3 evidence is presented that: Other mutants |
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Details of the target gene | |||||||||||||||||||||||||||
Gene Model of Rodent Parasite | PBANKA_1342500 | ||||||||||||||||||||||||||
Gene Model P. falciparum ortholog | PF3D7_1327300 | ||||||||||||||||||||||||||
Gene product | conserved Plasmodium protein, unknown function | ||||||||||||||||||||||||||
Gene product: Alternative name | SPM3 | ||||||||||||||||||||||||||
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Details of the genetic modification | |||||||||||||||||||||||||||
Name of the tag | GFP | ||||||||||||||||||||||||||
Details of tagging | C-terminal | ||||||||||||||||||||||||||
Additional remarks: tagging | |||||||||||||||||||||||||||
Commercial source of tag-antibodies | |||||||||||||||||||||||||||
Type of plasmid/construct | (Linear) plasmid single 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 endogenous tagging of PbSPM3 (PBANKA_1342500), a 992 bp long 3’ homology region was amplified from PbANKA wild-type genomic DNA. The reverse primer encoded the same linker sequence that was used for GFP-tagging of PfSPM3 as described in Birnbaum et al. 2017. The amplicon was cloned into the pL18 489 vector using EcoRI/BamHI restriction sites. Prior to transfection, the vector was linearized using the SwaI restriction enzyme followed by ethanol-precipitation | ||||||||||||||||||||||||||
Additional remarks selection procedure | |||||||||||||||||||||||||||
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