Back to search resultsSummaryRMgm-903
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Successful modification | The parasite was generated by the genetic modification |
The mutant contains the following genetic modification(s) | Gene tagging |
Reference (PubMed-PMID number) |
Reference 1 (PMID number) : 23638681 Reference 2 (PMID number) : 27084458 |
MR4 number | |
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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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The mutant parasite was generated 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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Name of the mutant parasite | |
RMgm number | RMgm-903 |
Principal name | PbDHHC9 _3xHA_TAG |
Alternative name | |
Standardized name | |
Is the mutant parasite cloned after genetic modification | Yes |
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Phenotype | |
Asexual blood stage | Schizont expression confirmed by IFA and Western Blotting. IFA co-localises the tagged protein with PbMSP1, in late, but not early schizonts, suggesting an IMC localisation. |
Gametocyte/Gamete | Not tested |
Fertilization and ookinete | Not tested |
Oocyst | DHHC3-HA, it was detectable in oocyst sporozoites, but not in salivary gland sporozoites. Generally the level of DHHC9-HA expression detectable by immunofluorescence was low in sporozoites. |
Sporozoite | DHHC3-HA, it was detectable in oocyst sporozoites, but not in salivary gland sporozoites. Generally the level of DHHC9-HA expression detectable by immunofluorescence was low in sporozoites. |
Liver stage | In liver stages, expression was not observed until 48 hours after invasion of the mammalian host cell, where it appeared to localize with dividing hepatic merozoites, a staining pattern that persisted into mature, 65 hour liver stage parasites. At 48 hours, DHHC9-HA was also localized to a discrete streak in the periphery of the parasitophorous vacuole, which showed partial overlap with staining for uis4 (up-regulated in infective sporozoites gene 4). This distinct DHHC9-HA streak was observed in all infected hepatocytes at this time point in liver stage development, but was absent from parasites at 65 hours after infection. |
Additional remarks phenotype | Mutant/mutation/phenotype Schizont expression confirmed by IFA and Western Blotting. IFA co-localises the tagged protein with PbMSP1, in late, but not early schizonts, suggesting an IMC localisation. 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 Reference PMID 27084458 |
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Details of the target gene | |||||||||||||||||||||||||||
Gene Model of Rodent Parasite | PBANKA_0932100 | ||||||||||||||||||||||||||
Gene Model P. falciparum ortholog | PF3D7_1115900 | ||||||||||||||||||||||||||
Gene product | palmitoyltransferase DHHC9 | ||||||||||||||||||||||||||
Gene product: Alternative name | DHHC9 | ||||||||||||||||||||||||||
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Details of the genetic modification | |||||||||||||||||||||||||||
Name of the tag | triple-HA | ||||||||||||||||||||||||||
Details of tagging | C-terminal | ||||||||||||||||||||||||||
Additional remarks: tagging | |||||||||||||||||||||||||||
Commercial source of tag-antibodies | |||||||||||||||||||||||||||
Type of plasmid/construct | (Linear) plasmid double cross-over | ||||||||||||||||||||||||||
PlasmoGEM (Sanger) construct/vector used | Yes | ||||||||||||||||||||||||||
Name of PlasmoGEM construct/vector | PbGEM-121234 | ||||||||||||||||||||||||||
Modified PlasmoGEM construct/vector used | No | ||||||||||||||||||||||||||
Plasmid/construct map | |||||||||||||||||||||||||||
Plasmid/construct sequence | |||||||||||||||||||||||||||
Restriction sites to linearize plasmid | |||||||||||||||||||||||||||
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 | |||||||||||||||||||||||||||
Additional remarks selection procedure | |||||||||||||||||||||||||||
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