SummaryRMgm-5264
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Successful modification | The parasite was generated by the genetic modification |
The mutant contains the following genetic modification(s) | Gene disruption |
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-5264 |
Principal name | PbSPM3-KO |
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 | Not different from wild type |
Sporozoite | Normal sporozoite formation inside oocysts. Strongly reduced sporozoite numbers in salivary glands. Strongly reduced (aberrant) motility. |
Liver stage | Not different from wild type |
Additional remarks phenotype | Mutant/mutation Protein (function) Phenotype Analysis of a mutant expressing a C-terminal GFP-tagged version of SPM3 (RMgm-5265) showed expression in oocysts and sporozoites. In sporozoites, the fluorescent signal extended from the apical end towards the rear around the nucleus. Subsequent co-staining with an anti-tubulin antibody revealed co-localization of PbSPM3-GFP with tubulin (Figure 4A) in midgut and salivary gland sporozoites. C-terminal tagging of PbSPM3 with GFP did not affect parasite life cycle progression, as SPM3-GFP parasites showed normal midgut infection and midgut oocyst numbers as well as numbers of both midgut and salivary gland sporozoites comparable to wild-type. 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 | |||||||||||||||||||||||||
Inducable system used | No | ||||||||||||||||||||||||
Additional remarks inducable system | |||||||||||||||||||||||||
Type of plasmid/construct used | (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 | |||||||||||||||||||||||||
Partial or complete disruption of the gene | Complete | ||||||||||||||||||||||||
Additional remarks partial/complete disruption | |||||||||||||||||||||||||
Selectable marker used to select the mutant parasite | hdhfr/yfcu | ||||||||||||||||||||||||
Promoter of the selectable marker | eef1a | ||||||||||||||||||||||||
Selection (positive) procedure | pyrimethamine | ||||||||||||||||||||||||
Selection (negative) procedure | No | ||||||||||||||||||||||||
Additional remarks genetic modification | For gene deletion of PbSPM3 (PBANKA_1342500), both a 3’ and a 5’ homology region (750 and 717 bps) were amplified from PbANKA wild-type-genomic DNA and cloned into the Pb262 vector using HindIII/XhoI and EcoRI/EcoRV restriction 476 sites. The Pb262 vector contains the hDHFR gene that allows for positive selection using the drug pyrimethamine. Prior to transfection, the vector was linearized using SacII and PmeI restriction enzymes followed by ethanol-precipitation. | ||||||||||||||||||||||||
Additional remarks selection procedure | |||||||||||||||||||||||||
Primer information: Primers used for amplification of the target sequences
![]() Primer information: Primers used for amplification of the target sequences
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