SummaryRMgm-5266
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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) : 36403748 |
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 | Yuda M, Nishi T |
Name Group/Department | Department of Medical Zoology |
Name Institute | Mie University School of Medicine |
City | Mie, Tsu |
Country | Japan |
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Name of the mutant parasite | |
RMgm number | RMgm-5266 |
Principal name | B9(-) |
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 | Normal numbers of motile salivary gland sporozoites are formed. While 30 wild-type sporozoites were sufficient to establish infection in all rats, even 1 × 10 B9(-) sporozoites were insufficient to establish infection in all rats. |
Liver stage | B9(-) parasites formed abnormal liver stages (LS) and displayed increased cell traversal activity. The number of B9(-) LS parasites formed in HepG2 cells was 6% that of wild-type parasites at 24 and 48 h post-inoculation (hpi) of sporozoites. The average diameter of the B9(-) LS parasites decreased to approximately 80 % that of wild-type LSs at 48 hpi. There was a decreased number of sporozoites entering the hepatocytes by productive invasion and reciprocally, an increased number of sporozoites that continued migration through the hepatocytes. Evidence is presented that B9(-) LS parasites develop in the nucleus of hepatocytes. |
Additional remarks phenotype | Mutant/mutation B9(-) parasites formed abnormal liver stages (LS) and displayed increased cell traversal activity. The number of B9(-) LS parasites formed in HepG2 cells was 6% that of wild-type parasites at 24 and 48 h post-inoculation (hpi) of sporozoites. The average diameter of the B9(-) LS parasites decreased to approximately 80% that of wild-type LSs at 48 hpi. There was a decreased number of sporozoites entering the hepatocytes by productive invasion and reciprocally, an increased number of sporozoites that continued migration through the hepatocytes. Evidence is presented that B9(-) LS parasites develop in the nucleus of hepatocytes. Evidence is presented that sporozoites lose the capacity to switch invasion modes by disruption of either B9 or P52 (see also mutants RMgm-5267 and RMgm-5268). Additional information From the paper: We also performed cell wounding and membrane repair assays to evaluate the cell traversal activity of B9(-) sporozoites . The number of HepG2 cells damaged by invasion of B9(-) sporozoites was approximately 3 fold higher than that of wild type parasites. These results suggest that the disruption of B9 impaired the switching capacity of sporozoites . More specifically, there was a decreased number of sporozoites entering the hepatocytes by productive invasion and reciprocally, an increased number of sporozoites that continued migration through the hepatocytes . This phenotype of B9(-). is similar to the previously reported phenotypes of P52 disrupted P52(-) and P36 disrupted P36(-) parasites |
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Details of the target gene | |||||||||||||||||||||||||
Gene Model of Rodent Parasite | PBANKA_0808100 | ||||||||||||||||||||||||
Gene Model P. falciparum ortholog | PF3D7_0317100 | ||||||||||||||||||||||||
Gene product | 6-cysteine protein | ||||||||||||||||||||||||
Gene product: Alternative name | B9 | ||||||||||||||||||||||||
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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 | pbdhfr | ||||||||||||||||||||||||
Promoter of the selectable marker | pbdhfr | ||||||||||||||||||||||||
Selection (positive) procedure | pyrimethamine | ||||||||||||||||||||||||
Selection (negative) procedure | No | ||||||||||||||||||||||||
Additional remarks genetic modification | B9(-) parasites were generated by the insertion of a pyrimethamine-resistant gene into the b9 gene locus of wild-type parasites by homologous recombination. | ||||||||||||||||||||||||
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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