Successful modification | The parasite was generated by the genetic modification |
The mutant contains the following genetic modification(s) |
Gene disruption
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Reference (PubMed-PMID number) |
Reference 1 (PMID number) : 23434509 |
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
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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 | Patzewitz EM; Tewari R |
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-840 |
Principal name | Δshlp1 |
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 | Strongly reduced (55-60%) ookinete production. The motility of ookinetes that are produced is normal. |
Oocyst | Strongly reduced (55-60%) ookinete production. No oocysts are formed. |
Sporozoite | Not tested |
Liver stage | Not tested |
Additional remarks phenotype | Mutant/mutation
The mutant lacks expression of shewanella-like protein phosphatase 1 (SHLP1).
Protein (function)
Bioinformatic analysis of the Plasmodium genome has revealed the presence of non-conventional protein phosphatases (PPs) containing kelch-like motifs as found in plant PPs and two bacterial Shewanella-like PPs (Shelphs or SHLP), both lacking orthologs in humans. SHLPs are related to the phosphoprotein phosphatase (PPP) family of serine/threonine PPs (STPs), span the eukaryote-prokaryote boundary, and are also present in plants, heterokonts, fungi, and some Protozoa.
Phenotype
Strongly reduced (55-60%) ookinete production. Evidence is presented that this reduction is not due parasite death or dedifferentiation of ookinetes before they reach full maturity at 24 hr. This suggests that SHLP1 plays an important (though not essential) role at an early stage in ookinete development and differentiation.
The motility of ookinetes that are produced is normal. No oocysts are formed. Evidence is presented for aberrant formation of micronemes in ookinetes.
Additional information
Analysis of a mutant expressing a GFP-tagged version of SHLP1 (RMgm-841) showed diffuse fluorescence staining in all parasite stages examined by microscopy with prominent staining in the asexual stages, the male gamete, and oocyst. Despite the presence of a predicted apicoplast targeting signal, the protein was found distributed non-uniformly throughout the parasite cytoplasm in all stages analyzed and co-localized with ER tracker Red, suggesting that SHLP1-GFP is located in the endoplasmic reticulum (ER).
Genetic crossing of mutant male gametes with wild type female gametes resulted in the formation of oocysts indicating normal fertility of mutant male gametes.
Other mutants
RMgm-841: a mutant expressing a GFP-tagged version of SHLP1. |