| Successful modification | The parasite was generated by the genetic modification |
| The mutant contains the following genetic modification(s) |
Gene tagging
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| Reference (PubMed-PMID number) |
Reference 1 (PMID number) : 23699412
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| 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 507cl1 (RMgm-7)
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| Other information parent line | P.berghei ANKA 507cl1 (RMgm-7) is a reference ANKA mutant line which expresses GFP under control of a constitutive promoter. This reference line does not contain a drug-selectable marker (PubMed: PMID: 16242190). |
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| The mutant parasite was generated by |
| Name PI/Researcher | Ning, J.; Billker, O.; Snell, W.J. |
| Name Group/Department | Department of Cell Biology |
| Name Institute | University of Texas Southwestern Medical School |
| City | Dallas, Texas |
| Country | US |
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| Name of the mutant parasite |
| RMgm number | RMgm-880 |
| Principal name | PbGEX1-HA |
| Alternative name | |
| Standardized name | |
| Is the mutant parasite cloned after genetic modification | Yes |
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| Phenotype |
| Asexual blood stage | No PbGEX1-HA expression in asexual blood stages |
| Gametocyte/Gamete | PbGEX1-HA was detected in a circular pattern surrounding the nucleus of the microgametocyte and macrogametocyte. Circumnuclear staining persisted to the ookinete stage. |
| Fertilization and ookinete | PbGEX1-HA was detected in a circular pattern surrounding the nucleus of the microgametocyte and macrogametocyte. Circumnuclear staining persisted to the ookinete stage. |
| Oocyst | Not tested |
| Sporozoite | Not tested |
| Liver stage | Not tested |
| Additional remarks phenotype | Mutant/mutation
The mutant expresses a C-terminal triple-HA-tagged version of GEX1. The endogenous gex1 gene has been tagged with the triple-HA tag.
Protein (function)
PbGEX1 is an ortholog of Chlamydomonas Cre06.g280600 (CrGEX1), a protein that functions in nuclear fusion. It is similar to an Arabidopsis gene, GEX1, previously shown to be involved in gametophyte development and embryogenesis.
CrGEX1 and PbGEX1 are members of a large, previously unrecognized family whose first-characterized member is KAR5 which is a Saccharomyces cerevisiae nuclear envelope protein, essential for nuclear fusion during yeast sexual reproduction
Phenotype
Phenotype analyses of a mutant lacking GEX1 expression (RMgm-879) indicate a defect of oocyst development. Oocysts either remained small and retained a compact nucleus or became highly vacuolated but invariably failed to show signs of sporogony. Normal numbers of ookinetes are produced. Evidence is presented that DNA synthesis/replication during meiosis in ookinetes is normal, resulting in 4N (tetraploid) ookinetes. Evidence is presented that an uncharacterized event in the nuclear envelope was altered in the mutant zygotes.
Analyses of the mutant expressing the HA-tagged version of GEX1 indicate the absence of GEX1 expression in asexual blood stages. PbGEX1-HA was detected in a circular pattern surrounding the nucleus of the microgametocyte and macrogametocyte. Circumnuclear staining persisted to the ookinete stage. The gex1/PbGEX1-HA progeny behaved as wild type, demonstrating that the tagged protein was functional. This and other data presented in the paper indicate that GEX1 is a nuclear envelope protein.
Other mutants
A mutant lacking GEX1 expression (RMgm-879) |
*RMgm-880
