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) : 23144823 |
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 |
Not applicable
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Other information parent line | |
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The mutant parasite was generated by |
Name PI/Researcher | S. Iwanaga; M. Yuda |
Name Group/Department | Department of Medical Zoology |
Name Institute | Mie University School of Medicine |
City | Tsu, Mie |
Country | Japan |
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Name of the mutant parasite |
RMgm number | RMgm-792 |
Principal name | AP2-L(-) |
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 | Not different from wild type |
Oocyst | Not different from wild type |
Sporozoite | Not different from wild type |
Liver stage | Normal sporozoite production. Gliding motility, cell traversal and hepatocyte invasion of sporozoites is normal. Infectivity of sporozoites in vivo is 10.000 lower compared to wild type sporozoites (as measured by prepatent period). Development of liver stages is affected (see 'Additional remarks phenotype'). |
Additional remarks phenotype | Mutant/mutation
The mutant lacks expression of AP2-L (transcription factor with AP2 domain(s))
Protein (function)
Apetala 2 (AP2)-family proteins are transcription factors that have DNA-binding domains of ,60 amino acids called AP2 domains. Recently, AP2 genes have been found in the genomes of Plasmodium parasites. In P. falciparum 27 AP2-family genes have been identified. Among these genes, 26 are conserved in the other Plasmodium species whose entire genomes have been sequenced. Each member of this family has 1 to 4 AP2 domains, and the amino acid sequences of these domains are highly conserved among Plasmodium orthologs.
AP2-L encodes a protein of 1272 amino acids with two AP2 domains. These domains are located near the C-terminus of the protein and are separated from each other by a short linker.
Phenotype
The phenotype analyses indicate that the lack of expression of AP2-L does not affect blood stage and mosquito stage development. Gliding motility, cell traversal and hepatocyte invasion of sporozoites is normal. Liver stage development is strongly reduced.
At 24 hpi, the difference in size between the wild-type and AP2-L(2) liver stages was still subtle, but it became clear at 36 hpi. After 36 hpi, the sizes of the AP2-L(2) LS parasites scarcely increased, whereas the wild-type parasites continued to grow until 48 hpi. In accordance with the growth of the LS parasites, arrest of nuclear division became clear at 36 hpi in the AP2-L(2) parasites. The appearance of the nuclei remained the same, and no AP2-L(2) LS parasites in the cytomere stage were formed in the later stage. These observations indicate that the development of AP2-L(2) parasites almost completely arrested at about 36 hpi, i.e., in the middle of the schizont stage, which caused a severe decrease in their ability to infect the liver
Additional information
Analyses of a mutant expressing a C-terminal tagged version of AP2-L showed expression and localisation of AP2-L in the nucleus of erythrocytic trophozoites, oocyst-derived sporozoites, salivary gland sporozoites and liver stages.
Other mutants
RMgm-793: A mutant expressing a C-terminal GFP-tagged version of AP2-L
RMgm-794: A mutant expressing a C-terminal tagged mCherry version of AP2-L and expressing GFP under a constitutive promoter (eef1a). |