Back to search resultsSummaryRMgm-896
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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 | 2 |
Reference (PubMed-PMID number) |
Reference 1 (PMID number) : 23638681 |
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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 2.34 |
Other information parent line | P. 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/Researcher | K. Frénal, J.Rayner, O. Billker, E. Bushell, D. Soldati-Favre |
Name Group/Department | Malaria Programme |
Name Institute | Wellcome Trust Sanger Insitute |
City | Cambridge Hinxton |
Country | UK |
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Details of the target gene | |||||||||||||||||||||||||
Gene Model of Rodent Parasite | PBANKA_1419700 | ||||||||||||||||||||||||
Gene Model P. falciparum ortholog | PF3D7_1321400 | ||||||||||||||||||||||||
Gene product | palmitoyltransferase DHHC8, putative | ||||||||||||||||||||||||
Gene product: Alternative name | DHHC8 | ||||||||||||||||||||||||
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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 | Yes | ||||||||||||||||||||||||
Name of PlasmoGEM construct/vector | PbGEM-225987 | ||||||||||||||||||||||||
Modified PlasmoGEM construct/vector used | No | ||||||||||||||||||||||||
Plasmid/construct map | |||||||||||||||||||||||||
Plasmid/construct sequence | |||||||||||||||||||||||||
Restriction sites to linearize plasmid | |||||||||||||||||||||||||
Partial or complete disruption of the gene | Partial | ||||||||||||||||||||||||
Additional remarks partial/complete disruption | |||||||||||||||||||||||||
Selectable marker used to select the mutant parasite | hdhfr/yfcu | ||||||||||||||||||||||||
Promoter of the selectable marker | eef1a | ||||||||||||||||||||||||
Selection (positive) procedure | pyrimethamine | ||||||||||||||||||||||||
Selection (negative) procedure | No | ||||||||||||||||||||||||
Additional remarks genetic modification | The unsuccessful attempts to generate of mutants with a disrupted gene indicate an essential role during asexual blood stage growth/multiplication. This protein was selected in a study aimed at determining the repertoire of DHHC-CRD S-acyl transferase protein family as putative candidates for protein S-acyl transferases (PATs). The genes encoding these proteins were targeted for deletion to analyse their essentiality for blood stages In addition, mutants were generated that express a HA-tagged version of the protein to identify its cellular location. Background: Protein palmitoylation in particular is a fundamental, dynamic, and widespread posttranslational mechanism that controls transport, properties and activity of proteins across eukaryotes. Unlike other irreversible lipid modifications such as myristoylation and prenylation, the addition of a 16-carbon saturated palmitate group to the sulfhydryl group of a cysteine to form a hydroxylamine-sensitive thioester linkage, is a reversible modification. This constitutes a fast and dynamic mechanism to spatiotemporally control protein function by impacting reversibly on protein trafficking, stability and clustering. While palmitoylation frequently facilitates membrane association of a soluble protein by the addition of a hydrophobic anchor, this modification also occurs on transmembrane proteins, involving other effects such as structural conformation changes, protein-protein interactions or the clustering to specific lipid domains leading, for example, to assembly of signalling complexes. Palmitoylation can control the affinity of a protein for lipid membranes, which allows it to impact protein trafficking, stability, folding, signalling and interactions. The publication of the palmitome of the schizont stage of P. falciparum implicated a role for palmitoylation in host cell invasion, protein export and organelle biogenesis. The enzymes mediating transfer of palmitate from palmitoyl-CoA to a protein substrate were first identified in Saccharomyces cerevisiae and subsequently in mammals. Protein S-acyl transferases (PATs) belong to the DHHC family of proteins that exhibit a catalytic Asp-His-His-Cys conserved motif located within a cysteine-rich domain (CRD) and frequently between two transmembrane regions, facing the cytosol. Substrate recognition and catalysis occur after the protein substrates have associated with membrane via another lipidation. Other mutants: Search the RMgm-databese for the text term DHHC-CRD for a number of other related mutants | ||||||||||||||||||||||||
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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