Back to search resultsSummaryRMgm-4184
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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 | Unknown |
Reference (PubMed-PMID number) |
Reference 1 (PMID number) : 28676844 |
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Parent parasite used to introduce the genetic modification | |
Rodent Malaria Parasite | P. yoelii |
Parent strain/line | P. y. yoelii 17XNL |
Name parent line/clone | Not applicable |
Other information parent line | |
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Attempts to generate the mutant parasite were performed by | |
Name PI/Researcher | Hart RJ; Aly ASI |
Name Group/Department | Department of Tropical Medicine |
Name Institute | Tulane University |
City | New Orleans |
Country | US |
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Details of the target gene | |||||||||||||||||||||||||
Gene Model of Rodent Parasite | PY17X_0805100 | ||||||||||||||||||||||||
Gene Model P. falciparum ortholog | PF3D7_0704700 | ||||||||||||||||||||||||
Gene product | phosphopantetheine adenylyltransferase, putative | ||||||||||||||||||||||||
Gene product: Alternative name | PPAT | ||||||||||||||||||||||||
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Details of the genetic modification | |||||||||||||||||||||||||
Inducable system used | No | ||||||||||||||||||||||||
Additional remarks inducable system | |||||||||||||||||||||||||
Type of plasmid/construct used | (Linear) 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 | hdhfr | ||||||||||||||||||||||||
Promoter of the selectable marker | eef1a | ||||||||||||||||||||||||
Selection (positive) procedure | pyrimethamine | ||||||||||||||||||||||||
Selection (negative) procedure | No | ||||||||||||||||||||||||
Additional remarks genetic modification | The unsuccessful attempts to disrupt this gene indicate an essential function during asexual blood stage development/multiplication. CoA is an essential cofactor for all prokaryotes and eukaryotes to support a large number of metabolic processes including fatty acid biosynthesis and oxidation, as well as carbohydrate and amino acid metabolism. In many living cells, the transport and utilization of pantothenic acid (vitamin B5, pantothenate in ionic form) is essential for CoA biosynthesis. The cellular machinery for the biosynthesis of CoA from exogenous pantothenate involves a putative pantothenate transporter (PAT) and five enzymes, PanK (Pantothenate Kinase), PPCS (Phosphopantothenylcysteine Synthase), PPCDC (Phosphopantothenylcysteine Decarboxylase), PPAT (Phosphopantetheine Adenylyltransferase), and DPCK (Dephospho-CoA Kinase). Previous studies showed that PAT, PanK1, and PanK2 genes are dispensable for blood stage development in mice but are crucial for oocyst development and sporozoite formation in the mosquito. In this study the following was shown: 'The intermediate enzymes PPCS (Phosphopantothenylcysteine Synthase), PPCDC (Phosphopantothenylcysteine Decarboxylase; RMgm-4183) were shown to be dispensable for asexual and sexual blood stage development, but they were essential for oocyst development and the production of sporozoites. However, the last two enzymes of this pathway, PPAT (Phosphopantetheine Adenylyltransferase; RMgm-4184) and DPCK (Dephospho-CoA Kinase; RMgm-4185), were essential for blood stage development. These results indicate alternative first substrate requirement for the malaria parasite, other than the canonical pantothenate, for the synthesis of CoA in the blood but not inside the mosquito midgut. Collectively, these studies indicate that CoA de novo biosynthesis is essential for both blood and mosquito stages'. | ||||||||||||||||||||||||
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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