RMgmDB - Rodent Malaria genetically modified Parasites
SummaryRMgm-4663
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Last modified: 8 August 2019, 15:01
*RMgm-4663| Successful modification | The 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 |
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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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| Attempts to generate the mutant parasite were performed by | |
| Name PI/Researcher | Wall RJ, Tewari R |
| Name Group/Department | School of Life Sciences, Queens Medical Centre |
| Name Institute | University of Nottingham |
| City | Nottingham |
| Country | London |
Disrupted: Mutant parasite with a disrupted gene| top of page | |||||||||||||||||||||||||
| Details of the target gene | |||||||||||||||||||||||||
| Gene Model of Rodent Parasite | PBANKA_0908500 | ||||||||||||||||||||||||
| Gene Model P. falciparum ortholog | PF3D7_1140500 | ||||||||||||||||||||||||
| Gene product | myosin K | ||||||||||||||||||||||||
| Gene product: Alternative name | MyoK | ||||||||||||||||||||||||
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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 | The 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 | |||||||||||||||||||||||||
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Primer information: Primers used for amplification of the target sequences
 Primer information: Primers used for amplification of the target sequences
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