SummaryRMgm-246
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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 | 4 |
Reference (PubMed-PMID number) | Not published (yet) |
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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 | P. berghei ANKA cl15cy1 |
Other information parent line | A reference wild type clone from the ANKA strain of P. berghei (PubMed: PMID: 17406255). |
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Attempts to generate the mutant parasite were performed by | |
Name PI/Researcher | T. Pace, C.J. Janse |
Name Group/Department | Dipartimento di Malattie Infettive Parassitarie ed Immunomediate |
Name Institute | Istituto Superiore di Sanità |
City | Rome |
Country | Italy |
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Details of the target gene | |||||||||||||||||||||||||
Gene Model of Rodent Parasite | PBANKA_0819900 | ||||||||||||||||||||||||
Gene Model P. falciparum ortholog | PF3D7_0919000 | ||||||||||||||||||||||||
Gene product | nucleosome assembly protein | ||||||||||||||||||||||||
Gene product: Alternative name | NAPS, nucleosome assembly protein S; SET | ||||||||||||||||||||||||
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Details of the genetic modification | |||||||||||||||||||||||||
Inducable system used | No | ||||||||||||||||||||||||
Additional remarks inducable system | |||||||||||||||||||||||||
Type of plasmid/construct used | 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 NAPS/SET indicate an essential role of this protein during asexual blood stage development. Disruption experiments were performed in the Leiden Malaria Research Group (exp. 921, 922; pl1239) and in Istituto Superiore di Sanità (Rome, Dr. T. Pace) using the same construct. Plasmodium contains two nucleosome assembly proteins, termed NAPL and NAPS (NAPS (or SET): PF1093c nucleosome binding protein; PBANKA_081990; NAPL: PFL0185c Nucleosome binding protein 1; PBANKA_060270). NAPL is predominantly localized in the cytoplasm, whereas NAPS has mainly a nuclear location. Both NAPS and NAPL interact with histones but only NAPS is able to deposit histones onto DNA. See also RMgm-217 and RMgm-218 for mutants expressing GFP-tagged versions of NAPS/SET under the control of different promoters. Attempts to disrupt NAPL and NAPS of P. falciparum (Gill et al., 2009, J Biol Chem. 2009, Jan 27; Gill J. et al, 2010 Malar J) and NAPS of P. berghei (RMgm-246) were not successful, indicating an essential role during asexual blood stage development. | ||||||||||||||||||||||||
Additional remarks selection procedure | |||||||||||||||||||||||||
Primer information: Primers used for amplification of the target sequences
Primer information: Primers used for amplification of the target sequences
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