RMgmDB - Rodent Malaria genetically modified Parasites


Malaria parasiteP. berghei
Genetic modification not successful
DisruptedGene model (rodent): PBANKA_1344100; Gene model (P.falciparum): PF3D7_1329100; Gene product: myosin F (MyoF)
PhenotypeNo phenotype has been described
Last modified: 8 August 2019, 14:59
Successful modificationThe gene/parasite could not be changed/generated by the genetic modification.
The following genetic modifications were attempted Gene disruption
Number of attempts to introduce the genetic modification Unknown
Reference (PubMed-PMID number) Reference 1 (PMID number) : 31283102
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
Attempts to generate the mutant parasite were performed by
Name PI/ResearcherWall RJ, Tewari R
Name Group/DepartmentSchool of Life Sciences, Queens Medical Centre
Name InstituteUniversity of Nottingham

  Disrupted: Mutant parasite with a disrupted gene
Details of the target gene
Gene Model of Rodent Parasite PBANKA_1344100
Gene Model P. falciparum ortholog PF3D7_1329100
Gene productmyosin F
Gene product: Alternative nameMyoF
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 usedNo
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 parasitetgdhfr
Promoter of the selectable markerpbdhfr
Selection (positive) procedurepyrimethamine
Selection (negative) procedureNo
Additional remarks genetic modificationThe unsuccessful attempts to disrupt this gene indicate an essential function during asexual blood stage growth.

The myosin superfamily is comprised of molecular motors present during early eukaryotic cell evolution. In unicellular parasites, they perform a wide variety of cellular functions that require movement, including differentiation, host interactions, and cell invasion. The myosin molecule contains three main domains: the N‐terminal head domain, which hydrolyses ATP and binds actin filaments; the neck domain/lever arm, which has an α‐helical structure containing up to six IQ motifs; and a tail region, which is required for cargo binding. There are six P. berghei myosins, two of these (MyoA and MyoB) have no tail region, and the remainder have a tail, which in the case of MyoF contains five WD40 repeats.
Quantitative reverse transcription polymerase chain reaction (qRT‐PCR) analyses of blood stages, ookinetes and sporozoites revealed that the Class XIV myosins MyoA (PBANKA_0135570), MyoB (PBANKA_0110300), and MyoE (PBANKA_0112200) were strongly transcribed in the invasive stages with an abundance of MyoE transcript in developing merozoites within schizonts. MyoF (PBANKA_1344100) was transcribed at all stages and was second only to MyoA in abundance. In contrast, low levels of transcription of MyoK (PBANKA_0908500) and MyoJ (PBANKA_1444500) could be seen throughout these life cycle stages.
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