SummaryRMgm-4404
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Successful modification | The parasite was generated by the genetic modification |
The mutant contains the following genetic modification(s) | Gene disruption, Introduction of a transgene |
Reference (PubMed-PMID number) |
Reference 1 (PMID number) : 29241041 |
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 | M. Zhang; V. Nussenzweig; W.J. Sullivan |
Name Group/Department | Department of Pharmacology and Toxicology |
Name Institute | Indiana University School of Medicine |
City | Indianapolis |
Country | USA |
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Name of the mutant parasite | |
RMgm number | RMgm-4404 |
Principal name | PbeIK1(-) |
Alternative name | PbeIK1(-) ko |
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 tested |
Fertilization and ookinete | Not tested |
Oocyst | Not tested |
Sporozoite | Not tested |
Liver stage | Not tested |
Additional remarks phenotype | Mutant/mutation Three eIF2α kinases, IK1 (PF14_0423; eukaryotic initiation factor 2alpha kinase 1), IK2 (PfA0380w; PBANKA_020580; serine/threonine protein kinase, putative), and PK4 (PFF1370w; PBANKA_112690; protein kinase PK4), have been identified in Plasmodium (Fenell C et al. 2009. Malar J. 8, 99). |
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Details of the target gene | |||||||||||||||||||||||||
Gene Model of Rodent Parasite | PbANKA_1308400 | ||||||||||||||||||||||||
Gene Model P. falciparum ortholog | PF3D7_1444500 | ||||||||||||||||||||||||
Gene product | eukaryotic initiation factor 2alpha kinase 1 | ||||||||||||||||||||||||
Gene product: Alternative name | IK1; eIK1 | ||||||||||||||||||||||||
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Details of the genetic modification | |||||||||||||||||||||||||
Inducable system used | No | ||||||||||||||||||||||||
Additional remarks inducable system | |||||||||||||||||||||||||
Type of plasmid/construct used | (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 | |||||||||||||||||||||||||
Partial or complete disruption of the gene | Partial | ||||||||||||||||||||||||
Additional remarks partial/complete disruption |
PbeIK1 knockouts were generated using the same method that we previously described for generation of PbeIK2 (-) parasites (Zhang et al., 2010). The 30- terminal of PbeIK1 coding sequence in the genome (PbANKA_1308400, 4.5kb) is adjacent (less than 500 bp) to PbANKA_130830. We deleted the 50- terminus (2.5-kb) of the PbeIK1 coding sequence to avoid interference with the downstream gene. The 2.5-kb of the 50-portion of this gene encodes the entire eIF2a kinase catalytic domain (Zhang et al., 2010). The replacement plasmid pBC_PbeIK1KO contains a 1-kb DNA fragment encoding the 5’-UTR of PbeIK1, the GFP cassette, the mutated dihydrofolate reductase (DHFR) gene that confers resistance to pyrimethamine, and a 1-kb DNA fragment encoding the PbeIK1 CDS (position 2500bp – 3500bp). | ||||||||||||||||||||||||
Selectable marker used to select the mutant parasite | hdhfr | ||||||||||||||||||||||||
Promoter of the selectable marker | pbdhfr | ||||||||||||||||||||||||
Selection (positive) procedure | pyrimethamine | ||||||||||||||||||||||||
Selection (negative) procedure | No | ||||||||||||||||||||||||
Additional remarks genetic modification | PbeIK1 knockouts were generated using the same method that we previously described for generation of PbeIK2 (-) parasites (Zhang et al., 2010). The 30- terminal of PbeIK1 coding sequence in the genome (PbANKA_1308400, 4.5kb) is adjacent (less than 500 bp) to PbANKA_130830. We deleted the 50- terminus (2.5-kb) of the PbeIK1 coding sequence to avoid interference with the downstream gene. The 2.5-kb of the 50-portion of this gene encodes the entire eIF2a kinase catalytic domain (Zhang et al., 2010). The replacement plasmid pBC_PbeIK1KO contains a 1-kb DNA fragment encoding the 5’-UTR of PbeIK1, the GFP cassette, the mutated dihydrofolate reductase (DHFR) gene that confers resistance to pyrimethamine, and a 1-kb DNA fragment encoding the PbeIK1 CDS (position 2500bp – 3500bp). | ||||||||||||||||||||||||
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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Type and details of transgene | |||||||||||||||||||
Is the transgene Plasmodium derived | Transgene: not Plasmodium | ||||||||||||||||||
Transgene name | GFP | ||||||||||||||||||
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Details of the genetic modification | |||||||||||||||||||
Inducable system used | No | ||||||||||||||||||
Additional remarks inducable system | |||||||||||||||||||
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 | pbdhfr | ||||||||||||||||||
Selection (positive) procedure | pyrimethamine | ||||||||||||||||||
Selection (negative) procedure | No | ||||||||||||||||||
Additional remarks genetic modification | PbeIK1 knockouts were generated using the same method that we previously described for generation of PbeIK2 (-) parasites (Zhang et al., 2010). The 30- terminal of PbeIK1 coding sequence in the genome (PbANKA_1308400, 4.5kb) is adjacent (less than 500 bp) to PbANKA_130830. We deleted the 50- terminus (2.5-kb) of the PbeIK1 coding sequence to avoid interference with the downstream gene. The 2.5-kb of the 50-portion of this gene encodes the entire eIF2a kinase catalytic domain (Zhang et al., 2010). The replacement plasmid pBC_PbeIK1KO contains a 1-kb DNA fragment encoding the 5’-UTR of PbeIK1, the GFP cassette, the mutated dihydrofolate reductase (DHFR) gene that confers resistance to pyrimethamine, and a 1-kb DNA fragment encoding the PbeIK1 CDS (position 2500bp – 3500bp). | ||||||||||||||||||
Additional remarks selection procedure | |||||||||||||||||||
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Other details transgene | |||||||||||||||||||
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Promoter | |||||||||||||||||||
Gene Model of Parasite | PbANKA_1308400 | ||||||||||||||||||
Gene Model P. falciparum ortholog | PF3D7_1444500 | ||||||||||||||||||
Gene product | eukaryotic initiation factor 2alpha kinase 1 | ||||||||||||||||||
Gene product: Alternative name | IK1; eIK1 | ||||||||||||||||||
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3'-UTR | |||||||||||||||||||
Gene Model of Parasite | Not available | ||||||||||||||||||
Gene product | Not available | ||||||||||||||||||
Gene product: Alternative name | |||||||||||||||||||
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Insertion/Replacement locus | |||||||||||||||||||
Replacement / Insertion | Replacement locus | ||||||||||||||||||
Gene Model of Parasite | PbANKA_1308400 | ||||||||||||||||||
Gene product | eukaryotic initiation factor 2alpha kinase 1 | ||||||||||||||||||
Gene product: Alternative name | IK1; eIK1 | ||||||||||||||||||
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