RMgmDB - Rodent Malaria genetically modified Parasites

Summary

RMgm-4432
Malaria parasiteP. berghei
Genotype
Genetic modification not successful
DisruptedGene model (rodent): PBANKA_1123400; Gene model (P.falciparum): PF3D7_0624500; Gene product: anaphase-promoting complex subunit 11, putative (APC11)
PhenotypeNo phenotype has been described
Last modified: 9 April 2018, 18:38
  *RMgm-4432
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 3
Reference (PubMed-PMID number) Reference 1 (PMID number) : 29618731
Parent parasite used to introduce the genetic modification
Rodent Malaria ParasiteP. berghei
Parent strain/lineP. berghei ANKA
Name parent line/clone P. berghei ANKA 2.34
Other information parent lineP. berghei ANKA 2.34 is a cloned, gametocyte producer line of the ANKA strain (PubMed: PMID: 15137943).
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
CityNottingham
CountryUK

  Disrupted: Mutant parasite with a disrupted gene
Details of the target gene
Gene Model of Rodent Parasite PBANKA_1123400
Gene Model P. falciparum ortholog PF3D7_0624500
Gene productanaphase-promoting complex subunit 11, putative
Gene product: Alternative nameAPC11
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/multiplication.

One essential component that drives the cell cycle, and particularly mitosis, in many eukaryotic systems is the Anaphase Promoting Complex/Cyclosome (APC/C). The APC/C is a multi-subunit E3 ubiquitin ligase that promotes cell-cycle progression by covalently tagging regulators such as securin and cyclin B1 with ubiquitin leading to their proteolysis by the proteasome. The mammalian APC/C has 14 core components, and several key adaptor subunits, including cell division cycle protein 20 (CDC20) and the related CDH1. Intriguingly, only four APC/C components have been identified as coded by the Plasmodium genome: APC10, APC11 and APC3 (a tetratricopeptide repeat [TPR] containing subunit), along with CDC20.
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