SummaryRMgm-5521
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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) : 39541984 |
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 cl15cy1 |
Other information parent line | A reference wild type clone from the ANKA strain of P. berghei (PubMed: PMID: 17406255). |
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The mutant parasite was generated by | |
Name PI/Researcher | Sayers C, Billker O |
Name Group/Department | The Laboratory for Molecular Infection Medicine Sweden |
Name Institute | Umeå University |
City | Umeå |
Country | Sweden |
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Name of the mutant parasite | |
RMgm number | RMgm-5521 |
Principal name | sun1-KO |
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 | Sun1-KO microgametocytes formed exflagellation centres when activated in vitro at the same rate as wild type. Sun1-KO ookinetes contained only half the DNA of wild type ookinetes, probably because most microgametes failed to deliver DNA to the zygote. This was NOT the result of defective replication during gametogenesis, but mitotic spindles were missing from nuclei, suggesting SUN1 is important for the microgamete nucleus or its genome to connect to the axoneme. |
Fertilization and ookinete | Morphologically mature ookinetes are formed, but very few oocysts present on the midguts of infected mosquitoes. Sun1-KO ookinetes contained only half the DNA of wild type ookinetes, probably because most microgametes failed to deliver DNA to the zygote. This was NOT the result of defective replication during gametogenesis, but mitotic spindles were missing from nuclei, suggesting SUN1 is important for the microgamete nucleus or its genome to connect to the axoneme. |
Oocyst | Strongly reduced oocyst formation. Morphologically mature ookinetes are formed, but very few oocysts present on the midguts of infected mosquitoes |
Sporozoite | Not tested |
Liver stage | Not tested |
Additional remarks phenotype | Mutant/mutation Evidence is presented for the following: |
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Details of the target gene | |||||||||||||||||||||||||
Gene Model of Rodent Parasite | PBANKA_1430900 | ||||||||||||||||||||||||
Gene Model P. falciparum ortholog | PF3D7_1215100 | ||||||||||||||||||||||||
Gene product | SUN domain-containing protein, putative | ||||||||||||||||||||||||
Gene product: Alternative name | SUN1 | ||||||||||||||||||||||||
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Details of the genetic modification | |||||||||||||||||||||||||
Inducable system used | No | ||||||||||||||||||||||||
Additional remarks inducable system | |||||||||||||||||||||||||
Type of plasmid/construct used | (Linear) PCR construct double cross-over | ||||||||||||||||||||||||
PlasmoGEM (Sanger) construct/vector used | Yes | ||||||||||||||||||||||||
Name of PlasmoGEM construct/vector | - | ||||||||||||||||||||||||
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 | hdhfr/yfcu | ||||||||||||||||||||||||
Promoter of the selectable marker | eef1a | ||||||||||||||||||||||||
Selection (positive) procedure | pyrimethamine | ||||||||||||||||||||||||
Selection (negative) procedure | No | ||||||||||||||||||||||||
Additional remarks genetic modification | The PlasmoGEM PBANKA_1430900 (SUN1) KO and HA-tagging transfection vectors were individually transfected into the PbANKA Cl15 cy1 parasite background line. Pyrimethamine-selected parasites were genotyped by PCR. | ||||||||||||||||||||||||
Additional remarks selection procedure | |||||||||||||||||||||||||
Primer information: Primers used for amplification of the target sequences
![]() Primer information: Primers used for amplification of the target sequences
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