RMgmDB - Rodent Malaria genetically modified Parasites
SummaryRMgm-190
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Last modified: 6 March 2009, 21:25
*RMgm-190| 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) : 18551176 |
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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 cl15cy1 |
| Other information parent line | A reference wild type clone from the ANKA strain of P. berghei (PubMed: PMID: 17406255). |
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| Attempts to generate the mutant parasite were performed by | |
| Name PI/Researcher | C. van Ooij; C.J. Janse; K. Haldar |
| Name Group/Department | Department of Pathology |
| Name Institute | Northwestern University |
| City | Chicago, Illinois |
| Country | USA |
Disrupted: Mutant parasite with a disrupted gene| top of page | |||||||||||||||||||||||||
| Details of the target gene | |||||||||||||||||||||||||
| Gene Model of Rodent Parasite | PBANKA_1327100 | ||||||||||||||||||||||||
| Gene Model P. falciparum ortholog | PF3D7_1463900 | ||||||||||||||||||||||||
| Gene product | EF-hand calcium-binding domain-containing protein, putative | ||||||||||||||||||||||||
| Gene product: Alternative name | |||||||||||||||||||||||||
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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 | No | ||||||||||||||||||||||||
| Modified PlasmoGEM construct/vector used | No | ||||||||||||||||||||||||
| Plasmid/construct map | |||||||||||||||||||||||||
| Plasmid/construct sequence | |||||||||||||||||||||||||
| Restriction sites to linearize plasmid | KpnI/SacII | ||||||||||||||||||||||||
| Partial or complete disruption of the gene | Complete | ||||||||||||||||||||||||
| Additional remarks partial/complete disruption | |||||||||||||||||||||||||
| Selectable marker used to select the mutant parasite | tgdhfr | ||||||||||||||||||||||||
| Promoter of the selectable marker | pbdhfr | ||||||||||||||||||||||||
| Selection (positive) procedure | pyrimethamine | ||||||||||||||||||||||||
| Selection (negative) procedure | No | ||||||||||||||||||||||||
| Additional remarks genetic modification | The gene has been identified in a study to identify proteins that are exported from the blood stage parasite to the host erythrocyte and that are conserved between P. falciparum and P. berghei. These proteins contain signal sequences and the Host-Targeting (HT) motif (van Ooij et al., 2008, PloS Pathogens 4, e10000084). Eleven conserved proteins were identified and 9 genes encoding these proteins were targeted for gene disruption in P. berghei. All attempts to disrupt these genes were unsuccessful, indication the essential nature of their proteins for blood stage development. Name of the transfection attempts experiments (Leiden Malaria Research Group: 719.1 Independent unsuccessful attempts to disrupt this gene has been reported (see RMgm103). The reverse primer for amplification of the 3'target region as presented in the paper is identical to the primer for another gene in tabel S7 of the supplementary material. Blast searches verified that this primer is not correct | ||||||||||||||||||||||||
| 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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