| Successful modification | The parasite was generated by the genetic modification |
| The mutant contains the following genetic modification(s) |
Gene tagging
|
| Reference (PubMed-PMID number) |
Reference 1 (PMID number) : 19932717
|
| MR4 number |
|
| top of page |
| 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). |
| top of page |
| 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 |
| top of page |
| Name of the mutant parasite |
| RMgm number | RMgm-355 |
| Principal name | PbLAP2/EGFP |
| Alternative name | |
| Standardized name | |
| Is the mutant parasite cloned after genetic modification | Yes |
| top of page |
| Phenotype |
| Asexual blood stage | Not different from wild type |
| Gametocyte/Gamete | Expression of the GFP-tagged LAP2 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 LAP2 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 CCp1 (LAP2; LCCL/lectin adhesive-like protein 2). The endogenous lap2 gene is tagged with gfp using a construct that integrates via a single cross-over recombination event, resulting in a gfp-tagged lap2 gene and a partial, non-functional lap2 copy.
Protein (function)
The lap2 (ccp1) 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 LAP2 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 LAP2/CCp1 (RMgm-118, RMgm-121, RMgm-124) indicate a role of this protein during oocyst development and sporozoite formation/production
The expression in gametocytes and ookinetes and the association of LAP2/CCp1 with the crystalloids is comparable to the expression and localisation pattern of two other LCCL domain-containing proteins, LAP1/CCp3/PbSR (RMgm-116) and LAP3/CCp5 (RMgm-356).
Additional information
Other mutants
RMgm-118: A mutant lacking expression of LAP2/CCp1
RMgm-121: A mutant lacking expression of LAP2/CCp1
RMgm-122: A mutant expressing GFP under the control of the promoter LAP2/CCp1
RMgm-124: A mutant lacking expression of both LAP2/CCP1 and PbSR/PSLAP/LAP1/CCp3
|
*RMgm-355
