SummaryRMgm-863
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Successful modification | The parasite was generated by the genetic modification |
The mutant contains the following genetic modification(s) | Gene disruption |
Reference (PubMed-PMID number) |
Reference 1 (PMID number) : 23634205 |
MR4 number | |
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Parent parasite used to introduce the genetic modification | |
Rodent Malaria Parasite | P. yoelii |
Parent strain/line | P. y. yoelii YM |
Name parent line/clone | Not applicable |
Other information parent line | |
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The mutant parasite was generated by | |
Name PI/Researcher | Huang, X; Preiser, PR |
Name Group/Department | Division of Molecular Genetics & Cell Biology |
Name Institute | School of Biological Sciences, Nanyang Technological University |
City | Singapore |
Country | Singapore |
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Name of the mutant parasite | |
RMgm number | RMgm-863 |
Principal name | SERA2-ko |
Alternative name | |
Standardized name | |
Is the mutant parasite cloned after genetic modification | Yes |
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Phenotype | |
Asexual blood stage | Growth, multiplication and virulence of blood stages were (slightly but significantly) reduced compared wild type (WT) parasites. |
Gametocyte/Gamete | Not tested |
Fertilization and ookinete | Not tested |
Oocyst | Not tested |
Sporozoite | Not tested |
Liver stage | Not tested |
Additional remarks phenotype | Mutant/mutation Protein (function) SERA genes of P. berghei and P. yoelii are arranged in a tandem cluster that contains two serine-type SERAs (PbSERA1, -2) and three cysteine-type SERAs (PbSERA3, -4 and -5). Sequence homology and syntenic organisation indicate that PySERA3 (PY000293), -4 (PY02062) and -5 (PY02063) are orthologs of P. falciparum SERA6, (PF3D7_0207500) -7 (PF3D7_0207400), -8 (PF3D7_0207300) Evidence has been presented that transcription of sera1 and sera2 are upregulated in the lethal YM strain compared to the non-lethal YA strain of P. yoelii. Analyses of mutants expressing GFP-tagged SERA1 (RMgm-864) and SERA2 (RMgm-865) it appears that both PySERA1 and 2 are expressed in the later stage of parasite maturation and that the proteases are located insight the parasitophorous vacuole (PV). However, it cannot be ruled out that the observed location in the PV is either due to the GFP tag preventing proper export or due to the fact that SERA are processed at the C-terminal end prior to activation which could result in the GFP tag remaining within the PV while the protease is exported further. However, no obvious free GFP was detected in the parasite schizont extract using anti-GFP antibody. P. berghei mutants: |
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Details of the target gene | |||||||||||||||||||||||||
Gene Model of Rodent Parasite | PY17X_0305600 | ||||||||||||||||||||||||
Gene Model P. falciparum ortholog | Not available | ||||||||||||||||||||||||
Gene product | Not available | ||||||||||||||||||||||||
Gene product: Alternative name | SERA2 | ||||||||||||||||||||||||
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Details of the genetic modification | |||||||||||||||||||||||||
Inducable system used | No | ||||||||||||||||||||||||
Additional remarks inducable system | |||||||||||||||||||||||||
Type of plasmid/construct used | Plasmid double 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 | |||||||||||||||||||||||||
Partial or complete disruption of the gene | Complete | ||||||||||||||||||||||||
Additional remarks partial/complete disruption | |||||||||||||||||||||||||
Selectable marker used to select the mutant parasite | tgdhfr | ||||||||||||||||||||||||
Promoter of the selectable marker | pbdhfr | ||||||||||||||||||||||||
Selection (positive) procedure | pyrimethamine | ||||||||||||||||||||||||
Selection (negative) procedure | No | ||||||||||||||||||||||||
Additional remarks genetic modification | No information is provided on the primer sequences used to PCR-amplify the targeting sequences | ||||||||||||||||||||||||
Additional remarks selection procedure | Transfected parasites were first selected through FACs sorting for the green fluorescent signal, and sorted parasites were subsequently diluted for cloning. | ||||||||||||||||||||||||
Primer information: Primers used for amplification of the target sequences
Primer information: Primers used for amplification of the target sequences
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