SummaryRMgm-4140
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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) : 28468674 |
MR4 number | |
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Parent parasite used to introduce the genetic modification | |
Rodent Malaria Parasite | P. chabaudi |
Parent strain/line | P. c.chabaudi AS |
Name parent line/clone | Not applicable |
Other information parent line | Cloned lines of Plasmodium chabaudi chabaudi AS were originally obtained from David Walliker, University of Edinburgh, UK and subsequently passaged through mice by injection of infected red blood cells (iRBC) at the MRC National Institute for Medical Research, UK |
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The mutant parasite was generated by | |
Name PI/Researcher | Cunningham DA; Langhorne J |
Name Group/Department | Malaria Immunology laboratory |
Name Institute | Francis Crick Institute |
City | London |
Country | UK |
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Name of the mutant parasite | |
RMgm number | RMgm-4140 |
Principal name | PcASΔsmacEFluc |
Alternative name | |
Standardized name | |
Is the mutant parasite cloned after genetic modification | Yes |
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Phenotype | |
Asexual blood stage | Evidence is presented that the absence of SMAC does not affect blood stage growth, course of parasitemia and tissue sequestration of Plasmodium chabaudi infected erythrocytes (see also RMgm-4139) |
Gametocyte/Gamete | Not tested |
Fertilization and ookinete | Not tested |
Oocyst | Not tested |
Sporozoite | Not tested |
Liver stage | Not tested |
Additional remarks phenotype | Mutant/mutation |
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Details of the target gene | |||||||||||||||||||||||||
Gene Model of Rodent Parasite | PCHAS_0101300 | ||||||||||||||||||||||||
Gene Model P. falciparum ortholog | Not available | ||||||||||||||||||||||||
Gene product | schizont membrane associated cytoadherence protein, putative | ||||||||||||||||||||||||
Gene product: Alternative name | SMAC | ||||||||||||||||||||||||
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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 | tgdhfr | ||||||||||||||||||||||||
Promoter of the selectable marker | pbdhfr | ||||||||||||||||||||||||
Selection (positive) procedure | pyrimethamine | ||||||||||||||||||||||||
Selection (negative) procedure | No | ||||||||||||||||||||||||
Additional remarks genetic modification | Plasmodium chabaudi AS parasites lacking expression of SMAC (PcASΔsmac; RMgm-4139) were generated by replacing the P. berghei smac (PBANKA_0100600) targeting region with the equivalent SMAC region of P. chabaudi (PCHAS_0101300; PCHAS_01_v3 52494-53200 and 53700-54284) using a published construct. Δsmac parasites expressing luciferase constitutively were subsequently generated using a modified construct (PcASΔsmacEFluc; see RMgm-4140) whereby eef1a luciferase was inserted within the targeting region at EcoRV/EcoR1 sites. Both Δsmac and ΔsmacEFluc constructs were digested with Kpn1/SacII prior to transfection. | ||||||||||||||||||||||||
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 | Luciferase (firefly) | ||||||||||||||||||
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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 | tgdhfr | ||||||||||||||||||
Promoter of the selectable marker | pbdhfr | ||||||||||||||||||
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_1133300 | ||||||||||||||||||
Gene Model P. falciparum ortholog | PF3D7_1357100 | ||||||||||||||||||
Gene product | elongation factor 1-alpha | ||||||||||||||||||
Gene product: Alternative name | eef1a | ||||||||||||||||||
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3'-UTR | |||||||||||||||||||
Gene Model of Parasite | Not available | ||||||||||||||||||
Gene product | Not available | ||||||||||||||||||
Gene product: Alternative name | |||||||||||||||||||
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Insertion/Replacement locus | |||||||||||||||||||
Replacement / Insertion | Replacement locus | ||||||||||||||||||
Gene Model of Parasite | PCHAS_0101300 | ||||||||||||||||||
Gene product | schizont membrane associated cytoadherence protein, putative | ||||||||||||||||||
Gene product: Alternative name | SMAC | ||||||||||||||||||
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