| 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) : 19932717
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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 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 | S. Saeed; V. Carter; J.T. Dessens |
| Name Group/Department | Department of Infectious and Tropical Diseases |
| Name Institute | London School of Hygiene & Tropical Medicine |
| City | London |
| Country | UK |
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| Name of the mutant parasite |
| RMgm number | RMgm-356 |
| Principal name | PbLAP3/EGFP |
| 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 | Expression of the GFP-tagged LAP3 protein in gametocytes. The protein was observed distributed throughout the parasite cytoplasm in a somewhat punctate pattern |
| Fertilization and ookinete | Expression of the GFP-tagged LAP3 protein in ookinetes. The protein was confined to two focal spots, often visibly associated with clusters of malaria pigment. Immunoelectronmicroscopy showed association of the protein with the crystalloids. |
| Oocyst | Not different from wild type |
| Sporozoite | Not different from wild type |
| Liver stage | Not tested |
| Additional remarks phenotype | Mutant/mutation
The mutant expresses a GFP-tagged version of LCCL domain-containing protein CCp5 (LAP3; LCCL/lectin adhesive-like protein 3). The endogenous lap3 gene is tagged with gfp using a construct that integrates via a double cross-over recombination event.
Protein (function)
The lap3 (ccp5) gene is a member of a small conserved gene family, encoding proteins with multiple adhesive domains, for example a Lgl1 (LCCL)-lectin adhesive domain. In P. falciparum the LCCL domain-containing proteins are termed PfCCp's and in P. berghei PbLAP's
Phenotype
Analysis of the location of the GFP-tagged LAP3 by fluorescence microscopy and immunoelectronmicroscopy shows that the protein is expressed in gametocytes and ookinetes and is associated with crystalloids in the ookinetes. Crystalloids are transient organelles that form in developing ookinetes and disappear after ookinete-to-oocyst transition. Plasmodium crystalloids are cytoplasmic aggregations of closely packed spherical particles 25–35 nm in diameter. In P. berghei, haemozoin-containing vacuoles accumulate around the edges of the crystalloid inclusion bodies.
Phenotype analyses of mutants lacking expression of different members of the LCCL domain-containing protein family (for example see LAP1/CCp3 RMgm-113, RMgm-114; LAP2/CCp1 RMgm-118, RMgm-121; LAP4/CCp2 RMgm-119; LAP6; CCp4 RMgm-120) indicate a role of several these proteins during oocyst development and sporozoite formation/production.
An unsuccessful attempt to disrupt LAP3/CCp5 has been described (RMgm-94)
The expression in gametocytes and ookinetes and the association of LAP3/CCp5 with the crystalloids is comparable to the expression and localisation pattern of two other LCCL domain-containing proteins, LAP1/CCp3/PbSR (RMgm-116) and LAP2/CCp1 (RMgm-355).
Additional information
Other mutants
RMgm-1249: The mutant expresses a GFP-tagged version of an mutated form of LAP3. The entire LCCL domain of the endogenous lap3 gene is removed.
RMgm-1250: A mutant lacking expression of LAP3 |
*RMgm-356
