RMgmDB - Rodent Malaria genetically modified Parasites
SummaryRMgm-390
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Last modified: 23 January 2011, 11:28
*RMgm-390| 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) : 20132450 |
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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 | K. Slavic; R. Tewari; S. Krishna |
| Name Group/Department | Centre for Infection, Cellular and Molecular Medicine |
| Name Institute | St. George's University of London |
| City | London |
| Country | UK |
Disrupted: Mutant parasite with a disrupted gene| top of page | |||||||||||||||||||||||||
| Details of the target gene | |||||||||||||||||||||||||
| Gene Model of Rodent Parasite | PBANKA_0302500 | ||||||||||||||||||||||||
| Gene Model P. falciparum ortholog | PF3D7_0204700 | ||||||||||||||||||||||||
| Gene product | hexose transporter | ||||||||||||||||||||||||
| Gene product: Alternative name | hexose transporter 1; PfHT1, PfHT, PbHT | ||||||||||||||||||||||||
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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, NotI | ||||||||||||||||||||||||
| 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 unsuccessful attempts to disrupt PbHT indicate an essential function during asexual blood stage growth (see also RMgm-603 for reports of unsuccessful attempts to disrupt pbht (pbht1)). In the same paper, unsuccessful attempts are reported to disrupt the hexose transporter (PfHT) in P. falciparum. PfHT is a facilitative hexose transporter in the Major Facilitator Superfamily of integral membrane proteins that mediates the uptake of glucose and fructose by the parasite. Pfht is a single copy gene in the P. falciparum genome with no close paralogues. Three other proteins, PFI0955w, PFI0785c and PFE1455w have been annotated with putative sugar transport function, although they have diverged considerably from typical sugar transporters (21%, 13% and 7% amino acid sequence identity compared with PfHT, respectively). Unlike PfHT, PFI0955w and PFI0785c are expressed only late in the asexual cycle. In the same paper it is reported that P. berghei parasites with a GFP-tagged version of PbHT were readily generated (see RMgm-391). This demonstrates that the locus is accessible for genetic targeting and confirms the essentiality of the hexose transporter gene for completion of the erythrocytic asexual cycle of malaria parasites. See also mutant RMgm-604 in which the endogenous hexose transporter 1 gene (pbht1) is replaced with the P. falciparum ortholog, pfht1. | ||||||||||||||||||||||||
| 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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