SummaryRMgm-5274
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*RMgm-5274| 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) : 39455824 |
| 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 17X |
| Name parent line/clone | Not applicable |
| Other information parent line | |
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| The mutant parasite was generated by | |
| Name PI/Researcher | Goswami D, Vaughan AM |
| Name Group/Department | Center for Global Infectious Disease Research |
| Name Institute | Seattle Children’s Research Institute |
| City | Seattle |
| Country | USA |
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| Name of the mutant parasite | |
| RMgm number | RMgm-5274 |
| Principal name | Py LINUP(mCherry) |
| 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 | LINUPmCherry expression was only seen during mid-to-late liver stage development, which in Py has an approximate 50-hour duration. LINUPmCherry expression was not detected at 24 hours of liver stage development but was detected at both 36 and 48 hours the latter a timepoint when merozoites form during the final stages of exo-erythrocytic schizogony. LINUPmCherry localized to live stage nuclei together with parasite DNA and partially co-localized with the histone marker, histone 3 acetylated lysine 9 (H3K9) which marks areas of active gene expression. |
| Additional remarks phenotype | Mutant/mutation Phenotype Analysis of the mutant expressing a C-terminal mCherry-tagged version of LINUP showed the following: LINUPmCherry expression was only seen during mid-to-late liver stage development, which in Py has an approximate 50-hour duration. LINUPmCherry expression was not detected at 24 hours of liver stage development but was detected at both 36 and 48 hours the latter a timepoint when merozoites form during the final stages of exo-erythrocytic schizogony. LINUPmCherry localized to live stage nuclei together with parasite DNA and partially co-localized with the histone marker, histone 3 acetylated lysine 9 (H3K9) which marks areas of active gene expression. Additional information |
Tagged: Mutant parasite with a tagged gene| top of page | |||||||||||||||||||||||||||
| Details of the target gene | |||||||||||||||||||||||||||
| Gene Model of Rodent Parasite | PY17X_1465200 | ||||||||||||||||||||||||||
| Gene Model P. falciparum ortholog | PF3D7_1249700 | ||||||||||||||||||||||||||
| Gene product | conserved Plasmodium protein, unknown function | ||||||||||||||||||||||||||
| Gene product: Alternative name | LINUP, liver stage nuclear protein | ||||||||||||||||||||||||||
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| Details of the genetic modification | |||||||||||||||||||||||||||
| Name of the tag | mCherry | ||||||||||||||||||||||||||
| Details of tagging | C-terminal | ||||||||||||||||||||||||||
| Additional remarks: tagging | |||||||||||||||||||||||||||
| Commercial source of tag-antibodies | |||||||||||||||||||||||||||
| Type of plasmid/construct | (Linear) 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 | |||||||||||||||||||||||||||
| Selectable marker used to select the mutant parasite | hdhfr | ||||||||||||||||||||||||||
| Promoter of the selectable marker | eef1a | ||||||||||||||||||||||||||
| Selection (positive) procedure | pyrimethamine | ||||||||||||||||||||||||||
| Selection (negative) procedure | No | ||||||||||||||||||||||||||
| Additional remarks genetic modification | Creation of Py LINUP(mCherry) utilized double crossover homologous recombination using modified plasmid pL0005 (obtained through the MR4 as part of the BEI Resources Repository, NIAID, NIH: Plasmodium berghei pL0005, MRA-774), which allowed for the addition of a mCherry epitope tag to the carboxy terminus of LINUP. | ||||||||||||||||||||||||||
| Additional remarks selection procedure | |||||||||||||||||||||||||||
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