Back to search resultsSummaryRMgm-4360
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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) : 28893907 |
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 |
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 | Green JL; Holder AA |
Name Group/Department | Malaria Parasitology Laboratory |
Name Institute | The Francis Crick Institute |
City | London |
Country | UK |
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Name of the mutant parasite | |
RMgm number | RMgm-4360 |
Principal name | MyoA-GFP |
Alternative name | |
Standardized name | |
Is the mutant parasite cloned after genetic modification | No |
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Phenotype | |
Asexual blood stage | Using live parasite microscopy, the protein was detected uniformly at the periphery of segmenting schizonts and extracellular merozoites in asexual blood stages and at a similar location in extracellular ookinetes and salivary gland sporozoites, consistent with location at the IMC. |
Gametocyte/Gamete | MyoA-GFP was not visible in either activated or non-activated gametocytes |
Fertilization and ookinete | Using live parasite microscopy, the protein was detected uniformly at the periphery of segmenting schizonts and extracellular merozoites in asexual blood stages and at a similar location in extracellular ookinetes and salivary gland sporozoites, consistent with location at the IMC. MyoA-GFP was not visible in either activated or non-activated gametocytes and barely detectable in the zygote and stage I and II ookinetes. Following the establishment of morphological polarity at about 10 hours after fertilization (Stage III) the protein was detectable in the growing protuberance rather than the spherical body in the intermediate retort forms, so-called because of their shape (stages III–V). This location was retained in later stages, with the MyoA-GFP associated with the parasite periphery in fully formed motile ookinetes. There was a concentration of fluorescence detected at the apical tip of the mature ookinete. |
Oocyst | Not tested |
Sporozoite | Using live parasite microscopy, the protein was detected uniformly at the periphery of segmenting schizonts and extracellular merozoites in asexual blood stages and at a similar location in extracellular ookinetes and salivary gland sporozoites, consistent with location at the IMC. In sporozoites there was also a clear perinuclear localization. |
Liver stage | In early cytomere stages of liver-stage schizonts the fluorescence signal appeared to be cytosolic, rather than membrane associated. In late cytomere stages, 55 h after invasion by sporozoites, a clear peripheral location associated with the hepatic merozoites was observed |
Additional remarks phenotype | Mutant/mutation
Using live parasite microscopy, the protein was detected uniformly at the periphery of segmenting schizonts and extracellular merozoites in asexual blood stages and at a similar location in extracellular ookinetes and salivary gland sporozoites, consistent with location at the IMC. MyoA-GFP was not visible in either activated or non-activated gametocytes and barely detectable in the zygote and stage I and II ookinetes. Following the establishment of morphological polarity at about 10 hours after fertilization (Stage III) the protein was detectable in the growing protuberance rather than the spherical body in the intermediate retort forms, so-called because of their shape (stages III–V). This location was retained in later stages, with the MyoA-GFP associated with the parasite periphery in fully formed motile ookinetes. There was a concentration of fluorescence detected at the apical tip of the mature ookinete. |
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Details of the target gene | |||||||||||||||||||||||||||
Gene Model of Rodent Parasite | PBANKA_1355700 | ||||||||||||||||||||||||||
Gene Model P. falciparum ortholog | PF3D7_1342600 | ||||||||||||||||||||||||||
Gene product | myosin A | ||||||||||||||||||||||||||
Gene product: Alternative name | MyoA; PfM-A | ||||||||||||||||||||||||||
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Details of the genetic modification | |||||||||||||||||||||||||||
Name of the tag | GFP | ||||||||||||||||||||||||||
Details of tagging | C-terminal | ||||||||||||||||||||||||||
Additional remarks: tagging | |||||||||||||||||||||||||||
Commercial source of tag-antibodies | |||||||||||||||||||||||||||
Type of plasmid/construct | (Linear) plasmid single cross-over | ||||||||||||||||||||||||||
PlasmoGEM (Sanger) construct/vector used | No | ||||||||||||||||||||||||||
Modified PlasmoGEM construct/vector used | No | ||||||||||||||||||||||||||
Plasmid/construct map | |||||||||||||||||||||||||||
Plasmid/construct sequence | |||||||||||||||||||||||||||
Restriction sites to linearize plasmid | |||||||||||||||||||||||||||
Selectable marker used to select the mutant parasite | hdhfr | ||||||||||||||||||||||||||
Promoter of the selectable marker | unknown | ||||||||||||||||||||||||||
Selection (positive) procedure | pyrimethamine | ||||||||||||||||||||||||||
Selection (negative) procedure | No | ||||||||||||||||||||||||||
Additional remarks genetic modification | |||||||||||||||||||||||||||
Additional remarks selection procedure | |||||||||||||||||||||||||||
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