SummaryRMgm-4647
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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) : 31251952 |
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 | Togiri J, 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-4647 |
Principal name | PbS23/SSP3 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 | Production of salivary gland sporozoites comparable to wild type. Normal sporozoite gliding motility, cell traversal and hepatocyte infection in vitro. |
Liver stage | Normal sporozoite gliding motility, cell traversal and hepatocyte infection in vitro. Normal growth (size) of liver stages in vitro at 12, 36 and 62 hour after infection of hepatocytes. Evidence is presented for a significant reduction in the formation of hepatic merozoites in PbS23/SSP3 KO mutants as revealed by DAPI staining. (Strongly) reduced infectivity in vivo (no blood infections or prolonged period after intravenous injection of sporozoites). |
Additional remarks phenotype | Mutant/mutation See also a P. yoelii mutant RMgm-1107 lacking expression of SSP3: this mutant shows a defect in sporozoite motility whereas no defect was detected in sporozoite infectivity |
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Details of the target gene | |||||||||||||||||||||||||
Gene Model of Rodent Parasite | PBANKA_1425200 | ||||||||||||||||||||||||
Gene Model P. falciparum ortholog | PF3D7_0812300 | ||||||||||||||||||||||||
Gene product | sporozoite surface protein 3, putative | ||||||||||||||||||||||||
Gene product: Alternative name | PbS23/SSP3 | ||||||||||||||||||||||||
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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 | eef1a | ||||||||||||||||||||||||
Selection (positive) procedure | pyrimethamine | ||||||||||||||||||||||||
Selection (negative) procedure | No | ||||||||||||||||||||||||
Additional remarks genetic modification | PbS23/SSP3 KO construct was generated by amplifying 573 bp for 5’ and 531 bp for 3’ fragments using primers FP6/RP6 and FP7/RP7 respectively. 5’ and 3’ fragments were cloned into pBC-GFP-hDHFR vector at XhoI/ClaI and NotI/AscI respectively. The targeting construct was released by XhoI and AscI digestion and gel purified. | ||||||||||||||||||||||||
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 | Insertion locus | ||||||||||||||||||
Gene Model of Parasite | PBANKA_1425200 | ||||||||||||||||||
Gene product | sporozoite surface protein 3, putative | ||||||||||||||||||
Gene product: Alternative name | PbS23/SSP3 | ||||||||||||||||||
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