RMgmDB - Rodent Malaria genetically modified Parasites
SummaryRMgm-483
|
||||||||||
Last modified: 21 December 2011, 16:26
*RMgm-483| 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) : 20584882 |
| top of page | |
| Parent parasite used to introduce the genetic modification | |
| Rodent Malaria Parasite | P. berghei |
| Parent strain/line | P. berghei ANKA |
| Name parent line/clone | Not applicable |
| Other information parent line | |
| top of page | |
| Attempts to generate the mutant parasite were performed by | |
| Name PI/Researcher | M. Zhang; V. Nussenzweig |
| Name Group/Department | Michael Heidelberger Division, Department of Pathology |
| Name Institute | New York University School of Medicine |
| City | New York |
| Country | USA |
Disrupted: Mutant parasite with a disrupted gene| top of page | |||||||||||||||||||||||||
| Details of the target gene | |||||||||||||||||||||||||
| Gene Model of Rodent Parasite | PBANKA_1126900 | ||||||||||||||||||||||||
| Gene Model P. falciparum ortholog | PF3D7_0628200 | ||||||||||||||||||||||||
| Gene product | protein kinase PK4 | eukaryotic translation initiation factor 2-alpha kinase | ||||||||||||||||||||||||
| Gene product: Alternative name | |||||||||||||||||||||||||
| top of page | |||||||||||||||||||||||||
| Details of the genetic modification | |||||||||||||||||||||||||
| Inducable system used | No | ||||||||||||||||||||||||
| Additional remarks inducable system | |||||||||||||||||||||||||
| Type of plasmid/construct used | Unknown | ||||||||||||||||||||||||
| 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 | Unknown | ||||||||||||||||||||||||
| Additional remarks partial/complete disruption | |||||||||||||||||||||||||
| Selectable marker used to select the mutant parasite | Unknown | ||||||||||||||||||||||||
| Promoter of the selectable marker | Unknown | ||||||||||||||||||||||||
| Selection (positive) procedure | Unknown | ||||||||||||||||||||||||
| Selection (negative) procedure | Unknown | ||||||||||||||||||||||||
| Additional remarks genetic modification | In the paper it is reported that multiple attempts to disrupt protein kinase PK4 in P. berghei were unsuccessful, indicating an essential role of PK4 during asexual blood stage development. No information/details is provided on the number of attempts to disrupt the gene, the construct used to disrupt the gene and the primers to amplify the target regions of pk4. One control mechanism of protein synthesis in eukaryotic cells involves the phosphorylation of the α-subunit of eukaryotic translation-initiation factor-2 (eIF-2α). During initiation of protein synthesis, a ternary complex of Met-tRNA, GTP, and the eukaryotic initiation factor 2 (eIF2) is delivered to the translation initiation machinery. During this translation process, eIF2-GTP is hydrolyzed to eIF2-GDP and released from the machinery. A guanine nucleotide exchange factor (eIF2B) facilitates the exchange of eIF2-GDP to eIF2-GTP, which is a requisite for a new round of translation. The phosphorylation of eIF2α blocks the recycling of eIF2-GTP and downregulates protein synthesis. The phosphorylation of eIF2α is associated with the appearance of stress granules in the cytoplasm of stressed cells. Three eIF2α kinases, IK1 (PF14_0423; eukaryotic initiation factor 2alpha kinase 1), IK2 (PfA0380w; PBANKA_020580; serine/threonine protein kinase, putative), and PK4 (PFF1370w; PBANKA_112690; protein kinase PK4), have been identified in Plasmodium. See RMgm-482 for a P. berghei mutant lacking expression of IK2 (PfA0380w; PBANKA_020580). Evidence has been presented that P. falciparum IK1 (PF14_0423; eukaryotic initiation factor 2alpha kinase 1) is not required for P. falciparum asexual and sexual blood stage development and sporozoite formation (Fenell C et al. 2009. Malar J. 8, 99) See also RMgm-566 for unsuccessful attempts to disrupt PBANKA_112690 Disruption of the P. falciparum ortholog has been attempted (Solyakov et al., 2011, Nat Commun, 2:565). The gene is likely essential for asexual proliferation as integration of the KO vector was not achieved. Accesibility of the locus to recombination was verified by C-terminal tagging. Disruption of the P. falciparum ortholog has been attempted (Solyakov et al., 2011, Nat Commun, 2:565). After transfection with a KO vector a strong PCR signal diagnostic for gene disruption was observed in transfected populations indicating that this gene is not essential for asexual proliferation. Cloning will however be required to validate this interpretation for this | ||||||||||||||||||||||||
| Additional remarks selection procedure | |||||||||||||||||||||||||
|
Primer information: Primers used for amplification of the target sequences
 Primer information: Primers used for amplification of the target sequences
| |||||||||||||||||||||||||
| top of page | |||||||||||||||||||||||||