SummaryRMgm-4288
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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) : 28708996 |
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 | PlasmoGEM (O. Billker; J. Rayner) |
Name Group/Department | PlasmoGEM |
Name Institute | Wellcome Trust Sanger Institute |
City | Hinxton Cambridge |
Country | UK |
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Name of the mutant parasite | |
RMgm number | RMgm-4288 |
Principal name | - |
Alternative name | |
Standardized name | |
Is the mutant parasite cloned after genetic modification | No |
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Phenotype | |
Asexual blood stage | The gene is dispensable. The growth rate not significantly different from wild type - [growth rate 1.03] - [95% CI: 0.98-1.09] - [PlasmoGem version 20170802] |
Gametocyte/Gamete | Not tested |
Fertilization and ookinete | Not tested |
Oocyst | Not tested |
Sporozoite | Not tested |
Liver stage | Not tested |
Additional remarks phenotype | Successful gene disruption as determined by barcode PCR in a large pool of gene-deletion mutants. P. berghei blood stages were transfected with a large pool of barcoded disruption (gene-deletion) vectors. These disruption vectors contained long modification arms to efficiently target the genes of interest. In addition the vectors contain gene-specific molecular barcodes. Co-transfecting multiple gene-deletion vectors in the same electroporation reproducibly generates complex pools of barcoded P. berghei mutants. Unsuccessful gene disruption/deletion or successful gene disruption/deletion is determined by the absence or presence of the barcode in the population (as determined by barcode-specific PCR analysis). This report is based on simultaneous phenotyping of mutants by barcode sequencing and is part of a large-scale genetic screen. The genotype of this mutant was not confirmed. The mutant was not isolated/cloned from the mixed population (large pool) of mutants.
The project website http://plasmogem.sanger.ac.uk/ has further information on methods and vector designs, provides tools for data visualisation and analysis, and allows researchers to request vectors to recreate mutants, confirm individual phenotypes and conduct more in-depth analyses. |
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Details of the target gene | |||||||||||||||||||||||||
Gene Model of Rodent Parasite | PBANKA_1353800 | ||||||||||||||||||||||||
Gene Model P. falciparum ortholog | PF3D7_1340400 | ||||||||||||||||||||||||
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 | Yes | ||||||||||||||||||||||||
Name of PlasmoGEM construct/vector | - | ||||||||||||||||||||||||
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 | P. berghei blood stages were transfected with a large pool of barcoded disruption (gene-deletion) vectors. These disruption vectors contained long modification arms to efficiently target the genes of interest. In addition the vectors contain gene-specific molecular barcodes. Co-transfecting multiple gene-deletion vectors in the same electroporation reproducibly generates complex pools of barcoded P. berghei mutants. Unsuccessful gene disruption/deletion or successful gene disruption/deletion is determined by the absence or presence of the barcode in the population (as determined by barcode-specific PCR analysis). | ||||||||||||||||||||||||
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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