SummaryRMgm-4072
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Successful modification | The parasite was generated by the genetic modification |
The mutant contains the following genetic modification(s) | Gene disruption, Introduction of a transgene |
Reference (PubMed-PMID number) |
Reference 1 (PMID number) : 28067322 |
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 | Al-Nihmi FM, Kumar KA |
Name Group/Department | Department of Animal Biology, School of Life Sciences |
Name Institute | University of Hyderabad |
City | Hyderabad |
Country | India |
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Name of the mutant parasite | |
RMgm number | RMgm-4072 |
Principal name | pbspeld 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 (salivary gland) sporozoite production. Normal gliding motility. Transmission of pbspeld ko sporozoites through mosquito bite did not initiate blood stage infection. High doses (2 × 104) of pbspeld ko sporozoites delivered through intravenously route led to occasional break through infection with a pre patent period of 9 days versus of 3.5 days for WT sporozoites |
Liver stage | Normal (salivary gland) sporozoite production. Normal gliding motility. Transmission of pbspeld ko sporozoites through mosquito bite did not initiate blood stage infection. High doses (2 × 104) of pbspeld ko sporozoites delivered through intravenously route led to occasional break through infection with a pre patent period of 9 days versus of 3.5 days for WT sporozoites. Normal hepatocyte invasion of mutant sporozoites. Mutants showed normal pattern of intracellular differentiation into EEFs at 12 hrs post infection of HepG2 cells and were similar in size to WT EEFs. At 36 hours the mutant EEFs were smaller than WT, and the difference in size was more pronounced at 62 hrs time point that revealed a distinct block in EEF development likely around 36 hr. |
Additional remarks phenotype | Mutant/mutation Phenotype |
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Details of the target gene | |||||||||||||||||||||||||
Gene Model of Rodent Parasite | PBANKA_0910900 | ||||||||||||||||||||||||
Gene Model P. falciparum ortholog | PF3D7_1137800 | ||||||||||||||||||||||||
Gene product | sporozoite surface protein essential for liver stage development, putative | ||||||||||||||||||||||||
Gene product: Alternative name | SPELD | ||||||||||||||||||||||||
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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 | ||||||||||||||||||||||||
Promoter of the selectable marker | unknown | ||||||||||||||||||||||||
Selection (positive) procedure | pyrimethamine | ||||||||||||||||||||||||
Selection (negative) procedure | No | ||||||||||||||||||||||||
Additional remarks genetic modification | |||||||||||||||||||||||||
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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Type and details of transgene | |||||||||||||||||||
Is the transgene Plasmodium derived | Transgene: not Plasmodium | ||||||||||||||||||
Transgene name | GFP | ||||||||||||||||||
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Details of the genetic modification | |||||||||||||||||||
Inducable system used | No | ||||||||||||||||||
Additional remarks inducable system | |||||||||||||||||||
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 | hdhfr | ||||||||||||||||||
Promoter of the selectable marker | eef1a | ||||||||||||||||||
Selection (positive) procedure | pyrimethamine | ||||||||||||||||||
Selection (negative) procedure | No | ||||||||||||||||||
Additional remarks genetic modification | |||||||||||||||||||
Additional remarks selection procedure | |||||||||||||||||||
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Other details transgene | |||||||||||||||||||
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Promoter | |||||||||||||||||||
Gene Model of Parasite | PBANKA_0711900 | ||||||||||||||||||
Gene Model P. falciparum ortholog | PF3D7_0818900 | ||||||||||||||||||
Gene product | heat shock protein 70 | ||||||||||||||||||
Gene product: Alternative name | HSP70 | ||||||||||||||||||
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3'-UTR | |||||||||||||||||||
Gene Model of Parasite | PBANKA_0711900 | ||||||||||||||||||
Gene product | heat shock protein 70 | ||||||||||||||||||
Gene product: Alternative name | HSP70 | ||||||||||||||||||
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Insertion/Replacement locus | |||||||||||||||||||
Replacement / Insertion | Replacement locus | ||||||||||||||||||
Gene Model of Parasite | PBANKA_0910900 | ||||||||||||||||||
Gene product | sporozoite surface protein essential for liver stage development, putative | ||||||||||||||||||
Gene product: Alternative name | SPELD | ||||||||||||||||||
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