Summary

RMgm-1804
Malaria parasiteP. berghei
Genotype
DisruptedGene model (rodent): PBANKA_0312600; Gene model (P.falciparum): PF3D7_0215900; Gene product: palmitoyltransferase DHHC11, putative
Phenotype Asexual bloodstage;
Last modified: 2 August 2017, 21:14
  *RMgm-1804
Successful modificationThe 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
Parent parasite used to introduce the genetic modification
Rodent Malaria ParasiteP. berghei
Parent strain/lineP. berghei ANKA
Name parent line/clone Not applicable
Other information parent line
The mutant parasite was generated by
Name PI/ResearcherPlasmoGEM (O. Billker; J. Rayner)
Name Group/DepartmentPlasmoGEM
Name InstituteWellcome Trust Sanger Institute
CityHinxton Cambridge
CountryUK
Name of the mutant parasite
RMgm numberRMgm-1804
Principal name-
Alternative name
Standardized name
Is the mutant parasite cloned after genetic modificationNo
Phenotype
Asexual blood stageThe gene is dispensable. The growth rate not significantly different from wild type - [growth rate 0.96] - [95% CI: 0.75-1.17] - [PlasmoGem version 20170802]
Gametocyte/GameteNot tested
Fertilization and ookineteNot tested
OocystNot tested
SporozoiteNot tested
Liver stageNot tested
Additional remarks phenotype

Successful gene disruption as determined by barcode PCR in a large pool of gene-deletion mutants.
Successful gene deletion indicates that the gene is not essential for asexual blood stage growth/multiplication.

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.

Asexual blood stage phenotype:
Growth rate phenotypes were obtained by counting barcodes on a next generation sequencer daily between days 4 and 8 post transfection. It is in the nature of the screen that genotypes of individual mutants were not validated.

  • Essential: growth rate not significantly different from 0.1.
  • Dispensable: growth rate not significantly different from 1 (corresponding to wild type).
  • Slow: growth rate significantly above 0.1 but below 1 (p < 0.05).
  • Fast: growth rate significantly above 1 (p < 0.05).
     

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.


  Disrupted: Mutant parasite with a disrupted gene
Details of the target gene
Gene Model of Rodent Parasite PBANKA_0312600
Gene Model P. falciparum ortholog PF3D7_0215900
Gene productpalmitoyltransferase DHHC11, putative
Gene product: Alternative name
Details of the genetic modification
Inducable system usedNo
Additional remarks inducable system
Type of plasmid/construct used(Linear) plasmid double cross-over
PlasmoGEM (Sanger) construct/vector usedYes
Name of PlasmoGEM construct/vector-
Modified PlasmoGEM construct/vector usedNo
Plasmid/construct map
Plasmid/construct sequence
Restriction sites to linearize plasmid
Partial or complete disruption of the geneComplete
Additional remarks partial/complete disruption
Selectable marker used to select the mutant parasitehdhfr/yfcu
Promoter of the selectable markereef1a
Selection (positive) procedurepyrimethamine
Selection (negative) procedureNo
Additional remarks genetic modificationP. 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  Click to view information
Primer information: Primers used for amplification of the target sequences  Click to hide information
Sequence Primer 1
Additional information primer 1
Sequence Primer 2
Additional information primer 2
Sequence Primer 3
Additional information primer 3
Sequence Primer 4
Additional information primer 4
Sequence Primer 5
Additional information primer 5
Sequence Primer 6
Additional information primer 6