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) : 28067322 |
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 | Al-Nihmi FM, Kumar KA |
Name Group/Department | Department of Animal Biology, School of Life Sciences |
Name Institute | University of Hyderabad |
City | Hyderabad |
Country | India |
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Name of the mutant parasite |
RMgm number | RMgm-4073 |
Principal name | pbspeldmCherry |
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 | At day 14, midgut oocysts, sporozoites within oocyst and at day 18–21, the salivary glands sporozoites were all positive for mCherry expression. Deconvoluted images of intact salivary glands harbouring sporozoites revealed the localization of mCherry to the sporozoite plasma membrane. The membrane associated mCherry colocalised with CSP in both midgut and salivary gland sporozoites. |
Sporozoite | At day 14, midgut oocysts, sporozoites within oocyst and at day 18–21, the salivary glands sporozoites were all positive for mCherry expression. Deconvoluted images of intact salivary glands harbouring sporozoites revealed the localization of mCherry to the sporozoite plasma membrane. The membrane associated mCherry colocalised with CSP in both midgut and salivary gland sporozoites. |
Liver stage | At day 14, midgut oocysts, sporozoites within oocyst and at day 18–21, the salivary glands sporozoites were all positive for mCherry expression. Deconvoluted images of intact salivary glands harbouring sporozoites revealed the localization of mCherry to the sporozoite plasma membrane. The membrane associated mCherry colocalised with CSP in both midgut and salivary gland sporozoites. The localization of mCherry was observed to the membrane of developing EEF at 17 hrs that colocalised with UIS4, a parasitophorous vacuolar membrane protein. However, no mcherry expression was associated with EEFs at 26, 36 and 62 hours post infection. |
Additional remarks phenotype | Mutant/mutation
The mutant expresses a C-terminal tagged mCherry version of SPELD
Protein (function)
SPELD was identified as the most abundantly expressed sequence tag in a sporozoite SAGE transcriptome analysis. Homologs of SPELD were not detected in other apicomplexa. PbSPELD sequence was analysed for the presence of transmembrane regions using multiple programs (TMHMM, TMpred, DAS, TopPred2), which predicted a single transmembrane region that is shared by all Plasmodium SPELD proteins. The SPELD proteins contain a tyrosine-rich domain, and the most abundant amino acid is tyrosine.
Phenotype analyses of a mutant lacking expression of SPELD (RMgm-4072) showed strongly reduced infectivity of sporozoites that was the result of aberrant EEF development
Phenotype
No expression in asexual stages,gametocytes, ookinetes and young oocysts.
At day 14, midgut oocysts, sporozoites within oocyst and at day 18–21, the salivary glands sporozoites were all positive for mCherry expression. Deconvoluted images of intact salivary glands harbouring sporozoites revealed the localization of mCherry to the sporozoite plasma membrane. The membrane associated mCherry colocalised with CSP in both midgut and salivary gland sporozoites. The localization of mCherry was observed to the membrane of developing EEF at 17 hrs that colocalised with UIS4, a parasitophorous vacuolar membrane protein. However, no mcherry expression was associated with EEFs at 26, 36 and 62 hours post infection.
Additional information
Other mutants
A mutant lacking expression of SPELD (RMgm-4072) |