SummaryRMgm-5526
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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) : 38195464 |
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 | Niikura M, Kobayashi F |
Name Group/Department | Department of Environmental Science, School of Life and Environmental Science |
Name Institute | Azabu University |
City | Kanagawa |
Country | Japan |
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Name of the mutant parasite | |
RMgm number | RMgm-5526 |
Principal name | Δ0519900 |
Alternative name | |
Standardized name | |
Is the mutant parasite cloned after genetic modification | Yes |
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Phenotype | |
Asexual blood stage | Mice infected with Δ0519900 mutants showed lower levels of parasitaemia than mice infected with control parasites and their survival was prolonged compared to mice infected with control parasites |
Gametocyte/Gamete | Not different from wild type |
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 | PBANKA_0519900 | ||||||||||||||||||||||||
Gene Model P. falciparum ortholog | PF3D7_1036900 | ||||||||||||||||||||||||
Gene product | conserved Plasmodium protein, unknown function | ||||||||||||||||||||||||
Gene product: Alternative name | |||||||||||||||||||||||||
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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 | To generated deletion mutants of P. berghei ANKA, the gene-targeting vectors for PBANKA_1019700, PBANKA_1101300 (sbp1) and PBANKA_0519900 were designed and constructed.Briefly, the 5′ and 3′ flanking regions of the open reading frame (ORF) of target genes were amplified by PCR. The PCR products were annealed to either side of the human dihydrofolate reductase (hdhfr)-expressing cassette, the red fluorescent protein gene (mCherry)-hdhfr-expressing cassette or the luciferase gene (luc2)-hdhfr-expressing cassette and amplified by PCR using gene-specific primers. The gene-targeting vectors were introduced into the 5′ and 3′ flanking regions of target genes by double-crossover homologous recombination. To generate transgenic parasites expressing mCherry-fused PBANKA_1019700, the gene-targeting vectors for PBANKA_1019700 were prepared by PCR. The PCR products were annealed to either side of the red fluorescent protein gene (mCherry)-hdhfr-expressing cassette and amplified by PCR using gene-specific primers. The gene-targeting vectors were introduced into the 3′ flanking regions of target genes by double-crossover homologous recombination. To generate transgenic parasites expressing GFP-fused PBANKA_0519900 and GFP-fused PBANKA_1359300, the gene-targeting vectors for PBANKA_0519900 and PBANKA_1359300 were prepared by PCR, respectively. The PCR products were annealed to either side of the green fluorescent protein gene (gfp)-mutated human deoxyhypusine synthase (hdhps)-expressing cassette and amplified by PCR using gene-specific primers. | ||||||||||||||||||||||||
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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