RMgmDB - Rodent Malaria genetically modified Parasites
SummaryRMgm-160
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Last modified: 24 February 2009, 16:02
*RMgm-160| 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) : 14678331 |
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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 | |
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| Attempts to generate the mutant parasite were performed by | |
| Name PI/Researcher | P. Uzureau; C.J. Janse; A.P. Waters; C. Braun Breton |
| Name Group/Department | Unité de Biologie des Interactions Hôte-Parasite |
| Name Institute | Institut Pasteur |
| City | Paris |
| Country | France |
Disrupted: Mutant parasite with a disrupted gene| top of page | |||||||||||||||||||||||||
| Details of the target gene | |||||||||||||||||||||||||
| Gene Model of Rodent Parasite | PBANKA_0911700 | ||||||||||||||||||||||||
| Gene Model P. falciparum ortholog | PF3D7_1136900 | ||||||||||||||||||||||||
| Gene product | subtilisin-like protease 2 | ||||||||||||||||||||||||
| Gene product: Alternative name | SUB2 | ||||||||||||||||||||||||
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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 | Partial | ||||||||||||||||||||||||
| Additional remarks partial/complete disruption | The construct was specifically designed to lead to the loss of 400 bp coding for part of the subtilisin catalytic domain including the active serine residue (see Supplementary material, Fig S2 in: Uzureau et al.,2004,Cell Microbiol. 6:65-78). | ||||||||||||||||||||||||
| 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 | SUB2 is a subtilisin-like serine protease. In Plasmodium three subtilisin-like proteases (SUB1-3) have been identified. SUB2 has been shown to be associated with dense granules and/or micronemes, the contents of which are released during erythrocyte invasion. It has been proposed to function as a 'sheddase', involved in the release of MSP1 and AMA1 during invasion (Harris et al., 2005, PloS Pathogens, 3, e29). The failure to disrupt the gene indicates an essential function in the blood stages. The generation of a mutant containing the endogenous sub2 gene tagged with c-myc (RMgm-161) demonstrates that the sub2 locus is accessible to genetic modification (Uzureau et al.,2004,Cell Microbiol. 6:65-78). See also GenBank accession numbers AJ242629 and AF145052 for the correct sequence of the gene encoding SUB2. | ||||||||||||||||||||||||
| 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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