Back to search resultsSummaryRMgm-846
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Successful modification | The parasite was generated by the genetic modification |
The mutant contains the following genetic modification(s) | Gene disruption |
Reference (PubMed-PMID number) |
Reference 1 (PMID number) : 23468629 |
MR4 number | |
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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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The mutant parasite was generated by | |
Name PI/Researcher | Guttery DS; Tewari R; Staines HM |
Name Group/Department | Centre for Genetics and Genomics, School of Biology, Queens Medical Centre |
Name Institute | University of Nottingham |
City | Nottingham |
Country | UK |
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Name of the mutant parasite | |
RMgm number | RMgm-846 |
Principal name | Δpbcax |
Alternative name | |
Standardized name | |
Is the mutant parasite cloned after genetic modification | Yes |
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Phenotype | |
Asexual blood stage | Not different from wild type |
Gametocyte/Gamete | Not different from wild type |
Fertilization and ookinete | Normal gametocyte production. Normal exflagellation of male gametocytes. No ookinetes are formed. Parasites failed to develop further from ‘round’ form zygotes into mature ookinetes |
Oocyst | Normal gametocyte production. Normal exflagellation of male gametocytes. No ookinetes are formed. No oocysts are formed. |
Sporozoite | Not tested |
Liver stage | Not tested |
Additional remarks phenotype | Mutant/mutation Analysis of P. falciparum PfCAX has demonstrated Ca2+/H+ exchange activity, an ability to exchange a limited range of other divalent cations, and a high transport capacity but low affinity (Km value of ,2 mM) for Ca2+, when expressed in Xenopus oocytes. In vivo studies characterising PfCAX were consistent with an atypical localisation to the inner mitochondrial membrane, and an atypical function, where the protein provides a pathway for removal of Ca2+ from this organelle back into the parasite cytosol. The studies on P. berghei PbCAX provided no evidence for a location or role in mitochondria. |
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Details of the target gene | |||||||||||||||||||||||||
Gene Model of Rodent Parasite | PBANKA_0102300 | ||||||||||||||||||||||||
Gene Model P. falciparum ortholog | PF3D7_0603500 | ||||||||||||||||||||||||
Gene product | cation/H+ antiporter | ||||||||||||||||||||||||
Gene product: Alternative name | CAH; pfCAX; pbCAX | ||||||||||||||||||||||||
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Details of the genetic modification | |||||||||||||||||||||||||
Inducable system used | No | ||||||||||||||||||||||||
Additional remarks inducable system | |||||||||||||||||||||||||
Type of plasmid/construct used | (Linear) 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 | ApaI, 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 pbcax knock-out vectors were constructed for a double cross-over homologous recombination in the pBS-DHFR plasmid that contains a Toxoplasma gondii dhfr/ts cassette conferring resistance to pyrimethamine. The knock-out construct was generated by inserting 507 bp of the pbcax 5' untranslated (UTR) region upstream and 497 bp of the pbcax 3' UTR region downstream of the dhfr cassette. The final knock-out construct was digested with ApaI and NotI to release the fragment for transfection. Transfection of P. berghei parasites with the knock-out construct was carried out in both wild-type parasites and in a line that constitutively expresses cytosolic GFP | ||||||||||||||||||||||||
Additional remarks selection procedure | |||||||||||||||||||||||||
Primer information: Primers used for amplification of the target sequences
Primer information: Primers used for amplification of the target sequences
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