RMgmDB - Rodent Malaria genetically modified Parasites
SummaryRMgm-275
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Last modified: 16 June 2009, 09:02
*RMgm-275| 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 | 4 |
| Reference (PubMed-PMID number) |
Reference 1 (PMID number) : 19346470 |
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| Parent parasite used to introduce the genetic modification | |
| Rodent Malaria Parasite | P. yoelii |
| Parent strain/line | P. y. yoelii 17X |
| Name parent line/clone | Not applicable |
| Other information parent line | Attempts to disrupt the gene were performed in both the 17X (lethal) and 17XNL (not lethal) strain of P. yoelii |
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| Attempts to generate the mutant parasite were performed by | |
| Name PI/Researcher | H. Otsuki; M. Torii |
| Name Group/Department | Department of Molecular Parasitology |
| Name Institute | Ehime University Graduate School of Medicine |
| City | Toon, Ehime |
| Country | Japan |
Disrupted: Mutant parasite with a disrupted gene| top of page | |||||||||||||||||||||||||
| Details of the target gene | |||||||||||||||||||||||||
| Gene Model of Rodent Parasite | PY17X_1337400 | ||||||||||||||||||||||||
| Gene Model P. falciparum ortholog | Not available | ||||||||||||||||||||||||
| Gene product | duffy receptor, beta form precursor | ||||||||||||||||||||||||
| Gene product: Alternative name | erythrocyte binding like protein, EBL; PyEBL | ||||||||||||||||||||||||
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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 | |||||||||||||||||||||||||
| 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 | pbdhfr | ||||||||||||||||||||||||
| Selection (positive) procedure | pyrimethamine | ||||||||||||||||||||||||
| Selection (negative) procedure | No | ||||||||||||||||||||||||
| Additional remarks genetic modification | See Otsuki et. al., PNAS 2009 106:7167-72 for detailed information about generation of the pR6Cyt-B12 construct which was used to create the pR6Cyt + 5U-B12 knock out construct (see supporting information). PY04764 is a gene encoding a type I integral transmembrane protein encoded by the ebl (erythrocyte-binding-like) gene family. Upon release from the micronemes, EBL proteins recognize erythrocyte receptors and initiate the formation of the tight junction. Plasmodium vivax uses an EBL orthologue, PvDBP (Pv110810), to recognize the Duffy antigen on the erythrocyte surface. P. falciparum has an expanded family of EBL proteins, for example EBA175 (PF07_0128), EBA181/JESEBL (PFA0125c), EBA140/BAEBL (MAL13P1.60), EBL1 (PFD1145c). EBL proteins possess 2 Cys-rich regions conserved among EBL orthologues. The N-terminal Cys-rich region named the DBL (Duffy-binding-like) domain or region 2 recognizes a specific erythrocyte surface receptor. The C-terminal Cys-rich region named the C-cys domain or region 6 is located adjacent to the transmembrane domain, and the number and location of Cys residues are well conserved among known Plasmodium species. Region 6 exhibits structural similarity to the KIX-binding domain of the coactivator CREB-binding protein and has been proposed to be a protein trafficking signal for transportation to the micronemes. See RMgm-276 and RMgm-277 for mutants ecoding mutated forms of the protein encoded by Pv110810. | ||||||||||||||||||||||||
| 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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