RMgmDB - Rodent Malaria genetically modified Parasites

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Summary

RMgm-266
Malaria parasiteP. berghei
Genotype
Genetic modification not successful
DisruptedGene model (rodent): PBANKA_0416700; Gene model (P.falciparum): PF3D7_0904700; Gene product: bacterial histone-like protein (PbHU)
PhenotypeNo phenotype has been described
Last modified: 24 April 2009, 20:13
  *RMgm-266
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) : 19358847
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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).
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Attempts to generate the mutant parasite were performed by
Name PI/ResearcherN. Sasaki, S. Sato
Name Group/DepartmentDivision of Parasitology
Name InstituteThe National Institute for Medical Research
CityLondon
CountryUK
  Disrupted: Mutant parasite with a disrupted gene
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Details of the target gene
Gene Model of Rodent Parasite PBANKA_0416700
Gene Model P. falciparum ortholog PF3D7_0904700
Gene productbacterial histone-like protein
Gene product: Alternative namePbHU
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Details of the genetic modification
Inducable system usedNo
Additional remarks inducable system
Type of plasmid/construct usedPlasmid 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 modificationIn the plastid, genomic DNA is assembled into a compact, highly organised structure, the nucleoid, like in bacteria from which the plastid has evolved. Bacterial nucleoid formation depends on a group of bacterial histone-like DNA binding proteins (BHLs). The HU protein, which binds DNA in a sequence non-specific manner and bends the bound DNA, is the most abundant BHL in the bacterial cell. The nuclear genome of P. falciparum encodes a HU homolog (PfHU) that is probably involved in DNA compaction in the plastid. PfHU is targeted exclusively to the parasite’s plastid and bound its natural target – the plastid DNA – sequence-independently. PfHU complements the lack of HU in Escherichia coli.
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 15’-ggtaccGCACATCTTTTGATTTTTTAGCCT-3’
Additional information primer 1PbHU-F1-KpnI
Sequence Primer 25’-aagcttTTTGTTCTCTTGTATTTTAAGATA-3’
Additional information primer 2PbHU-R1-HindIII
Sequence Primer 35’-gatatcTGCCTTTTATTGTAAAGAAAATAA-3’
Additional information primer 3PbHU-F2-EcoRV
Sequence Primer 45’-ggatccTACCTTTAATACCCATTTGCATA-3’
Additional information primer 4PbHU-R2-BamHI
Sequence Primer 5
Additional information primer 5
Sequence Primer 6
Additional information primer 6
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Conceptual design of the database: Dr. C.J. Janse (Leiden Malaria Research Group, Leiden, The Netherlands).
Technical design and realization: StudioDrie StudioDrie; S. Mededovic and A. van Wigcheren