RMgmDB - Rodent Malaria genetically modified Parasites
SummaryRMgm-5250
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Last modified: 23 September 2022, 14:54
*RMgm-5250| 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 | 3 |
| Reference (PubMed-PMID number) |
Reference 1 (PMID number) : 36040030 |
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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 | Ambekar, SV, Mair GR |
| Name Group/Department | Biomedical Sciences, |
| Name Institute | Iowa State University |
| City | Ames |
| Country | USA |
Disrupted: Mutant parasite with a disrupted gene| top of page | |||||||||||||||||||||||||
| Details of the target gene | |||||||||||||||||||||||||
| Gene Model of Rodent Parasite | PBANKA_0807900 | ||||||||||||||||||||||||
| Gene Model P. falciparum ortholog | PF3D7_0317300 | ||||||||||||||||||||||||
| Gene product | conserved Plasmodium protein, unknown function | ||||||||||||||||||||||||
| Gene product: Alternative name | NUP335 | ||||||||||||||||||||||||
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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 | unknown | ||||||||||||||||||||||||
| Promoter of the selectable marker | unknown | ||||||||||||||||||||||||
| Selection (positive) procedure | pyrimethamine | ||||||||||||||||||||||||
| Selection (negative) procedure | No | ||||||||||||||||||||||||
| Additional remarks genetic modification | The unsuccessful attempts to delete the gene indicate an essential role during asexual blood stage growth/multiplication In the paper different genes are identified that encode putative nucleoporins (NUPs). Two of the 5 novel, putative Nups are predicted to be essential for asexual life cycle progression from the large-scale PlasmoGEM forward mutagenesis screen in P. berghei while the remaining 3 (Nup335, Nup176, Nup269) were not targeted in this screen. In order to provide additional evidence for a critical role in parasite biology, we attempted to generate knockout lines for these 3 Nups using established methodologies Knockouts of Nup335, Nup176, and Nup269 failed repeatedly (n = 3 transfections for each gene), consistent with an important or essential role for these and all known Nups in the P. berghei blood stage in maintaining a functional NPC. | ||||||||||||||||||||||||
| Additional remarks selection procedure | |||||||||||||||||||||||||
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Primer information: Primers used for amplification of the target sequences
 Primer information: Primers used for amplification of the target sequences
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