SummaryRMgm-4977
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Successful modification | The parasite was generated by the genetic modification |
The mutant contains the following genetic modification(s) | Gene disruption, Gene disruption |
Reference (PubMed-PMID number) |
Reference 1 (PMID number) : 33667710 |
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 | Not applicable |
Other information parent line | |
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The mutant parasite was generated by | |
Name PI/Researcher | Soga A, Fukumoto S |
Name Group/Department | National Research Center for Protozoan Diseases |
Name Institute | Obihiro University of Agriculture and Veterinary Medicine |
City | Inada-cho, Obihiro, Hokkaido |
Country | Japan |
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Name of the mutant parasite | |
RMgm number | RMgm-4977 |
Principal name | tgloII KO/cgloII 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 | Not tested |
Fertilization and ookinete | Not tested |
Oocyst | Not tested |
Sporozoite | Not tested |
Liver stage | Liver stage development in vivo was analysed by measuring the prepatent period in mice infected intravenously with 10.000 sporozoites. Liver stage development in cultured hepatocytes was determined by real-time RT-PCR. A one-day delay in prepatent period was observed and a significant reduction of parasite numbers in cultured hepatocytes. |
Additional remarks phenotype | Mutant/mutation Phenotype |
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Details of the target gene | |||||||||||||||||||||||||
Gene Model of Rodent Parasite | PBANKA_0604400 | ||||||||||||||||||||||||
Gene Model P. falciparum ortholog | PF3D7_1205700 | ||||||||||||||||||||||||
Gene product | targeted glyoxalase II | ||||||||||||||||||||||||
Gene product: Alternative name | tGLO2 | ||||||||||||||||||||||||
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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 | Complete | ||||||||||||||||||||||||
Additional remarks partial/complete disruption | |||||||||||||||||||||||||
Selectable marker used to select the mutant parasite | puromycin-N-acetyltransferase (pac) | ||||||||||||||||||||||||
Promoter of the selectable marker | hsp70 | ||||||||||||||||||||||||
Selection (positive) procedure | puromycin in vitro | ||||||||||||||||||||||||
Selection (negative) procedure | No | ||||||||||||||||||||||||
Additional remarks genetic modification | See mutant RMgm-4180 and RMgm-4455 for the transfection and selection method (selection with pac or bsd in vitro). For the tgloII targeting vector, pBS/5tgoII-5E3DPacPB-3tgloII, recombination arms obtained by PCR were sequentially cloned into the pBluescript SK + plasmid pacPB expression cassette [19]. For the cgloII targeting vector, pBS/5cgoII-5E3DBsdPB-3cgloII, recombination arms obtained by PCR were sequentially cloned into the pBluescript SK + plasmid bsdPB expression cassette | ||||||||||||||||||||||||
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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Details of the target gene | |||||||||||||||||||||||||
Gene Model of Rodent Parasite | PBANKA_1004000 | ||||||||||||||||||||||||
Gene Model P. falciparum ortholog | PF3D7_0406400 | ||||||||||||||||||||||||
Gene product | cytosolic glyoxalase II | ||||||||||||||||||||||||
Gene product: Alternative name | cGLO2 | ||||||||||||||||||||||||
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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 | Complete | ||||||||||||||||||||||||
Additional remarks partial/complete disruption | |||||||||||||||||||||||||
Selectable marker used to select the mutant parasite | blasticidin S deaminase (bsd) | ||||||||||||||||||||||||
Promoter of the selectable marker | hsp70 | ||||||||||||||||||||||||
Selection (positive) procedure | blasticydin in vitro | ||||||||||||||||||||||||
Selection (negative) procedure | No | ||||||||||||||||||||||||
Additional remarks genetic modification | See mutant RMgm-4180 and RMgm-4455 for the transfection and selection method (selection with pac or bsd in vitro). For the tgloII targeting vector, pBS/5tgoII-5E3DPacPB-3tgloII, recombination arms obtained by PCR were sequentially cloned into the pBluescript SK + plasmid pacPB expression cassette [19]. For the cgloII targeting vector, pBS/5cgoII-5E3DBsdPB-3cgloII, recombination arms obtained by PCR were sequentially cloned into the pBluescript SK + plasmid bsdPB expression cassette | ||||||||||||||||||||||||
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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