RMgmDB - Rodent Malaria genetically modified Parasites

Summary

RMgm-1130
Malaria parasiteP. berghei
Genotype
Genetic modification not successful
DisruptedGene model (rodent): PBANKA_1205900; Gene model (P.falciparum): PF3D7_1007700; Gene product: AP2 domain transcription factor AP2-I (ApiAP2; PfAP2-T (expressed in the trophozoite stage))
PhenotypeNo phenotype has been described
Last modified: 28 September 2014, 20:09
  *RMgm-1130
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) Not published (yet)
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/ResearcherSantos, J; Llinas, M
Name Group/DepartmentBiochemistry and Molecular Biology
Name InstitutePenn State University
CityPennsylvania
CountryUS

  Disrupted: Mutant parasite with a disrupted gene
Details of the target gene
Gene Model of Rodent Parasite PBANKA_1205900
Gene Model P. falciparum ortholog PF3D7_1007700
Gene productAP2 domain transcription factor AP2-I
Gene product: Alternative nameApiAP2; PfAP2-T (expressed in the trophozoite stage)
Details of the genetic modification
Inducable system usedNo
Additional remarks inducable system
Type of plasmid/construct usednot known
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 geneUnknown
Additional remarks partial/complete disruption This mutant has been reported on the 2014 (25th) Annual Molecular Parasitology Meeting, Woodshole. In the abstract no data is provided on the details of the construct used in the attempts to knock-out this gene. When these unsuccessful attempts are published, additional information will be added to this mutant-entry.
Selectable marker used to select the mutant parasitenot known
Promoter of the selectable markernot known
Selection (positive) procedurenot known
Selection (negative) procedurenot known
Additional remarks genetic modificationThis mutant has been reported on the 2014 (25th) Annual Molecular Parasitology Meeting, Woodshole. In the abstract no data is provided on the details of the construct used in the attempts to knock-out this gene. When these unsuccessful attempts are published, additional information will be added to this mutant-entry.

The unsuccessful attempts to knock-out this gene indicates that the protein is essential for blood stage development/multiplication.

From the Abstract:
C-terminal GFP-tagged PfAP2-T is expressed in trophozoites and localizes to the nucleus. Chromatin immunoprecipitation experiments followed by high-throughput sequencing (ChIP-seq) with PfAP2-T-GFP parasites indicate that PfAP2-T binds upstream of a number of rhoptry genes in vivo, as well as glideosome associated proteins (GAPs) and merozoite surface proteins (MSPs). Furthermore, immunoprecipitation assays followed by mass spectrometry demonstrate that PfAP2-T interacts with the protein Bromodomain 1 (BDP1), which has also recently been shown to associate with invasion genes. Attempts to disrupt either pfap2-t in P. falciparum or its orthologue in P. berghei (pbanka_120590) were unsuccessful and we have been unable to regulate protein expression via the FKBP destabilization system.
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