RMgmDB - Rodent Malaria genetically modified Parasites
SummaryRMgm-668
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*RMgm-668| 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) : 21958024 |
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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 2.34 |
| Other information parent line | P. berghei ANKA 2.34 is a cloned, gametocyte producer line of the ANKA strain (PubMed: PMID: 15137943). |
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| The mutant parasite was generated by | |
| Name PI/Researcher | Talman A.M.; Sinden R.E. |
| Name Group/Department | Department of Life Sciences |
| Name Institute | Imperial College |
| City | London |
| Country | UK |
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| Name of the mutant parasite | |
| RMgm number | RMgm-668 |
| Principal name | GEST-GFP |
| 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 | Expression of GEST-GFP in male and female gametocytes with higher expression in females. GEST-GFP signals re-distribute upon activation of gametocytes (see 'Additional remarks phenotype'). |
| Fertilization and ookinete | Not tested |
| Oocyst | Not tested |
| Sporozoite | Expression of GEST-GFP in sporozoites (see 'Additional remarks phenotype'). |
| Liver stage | Not tested |
| Additional remarks phenotype | Mutant/mutation Phenotype Phenotype analyses of a mutant lacking expression of GEST (see mutant RMgm-667) indicates a role of GEST in the egress of male and female gametes from the erythrocyte. In addition, these analyses indicate that GEST plays a role in cell traversal/wounding of sporozoites. Additional information In sporozoites the GEST-GFP signals showed overlap with the distribution of a micronemal protein AMA-1. GEST-GFP signals were also observed in the trails left by motile sporozoites, indicating secretion of GEST by sporozoites. Other mutants |
Tagged: Mutant parasite with a tagged gene| top of page | |||||||||||||||||||||||||||
| Details of the target gene | |||||||||||||||||||||||||||
| Gene Model of Rodent Parasite | PBANKA_1312700 | ||||||||||||||||||||||||||
| Gene Model P. falciparum ortholog | PF3D7_1449000 | ||||||||||||||||||||||||||
| Gene product | gamete egress and sporozoite traversal protein, putative | ||||||||||||||||||||||||||
| Gene product: Alternative name | GEST, gamete egress and sporozoite traversal protein | ||||||||||||||||||||||||||
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| Details of the genetic modification | |||||||||||||||||||||||||||
| Name of the tag | GFP | ||||||||||||||||||||||||||
| Details of tagging | C-terminal | ||||||||||||||||||||||||||
| Additional remarks: tagging | |||||||||||||||||||||||||||
| Commercial source of tag-antibodies | Rabbit anti-GFP (ABCAM, USA) | ||||||||||||||||||||||||||
| Type of plasmid/construct | Plasmid single 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 | SacII, KpnI, HpaI | ||||||||||||||||||||||||||
| 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 | The GEST-GFP plasmid was generated by amplifying a 1 kb region of the most 3′ edge of the coding sequence of GEST with primers 10 and 14 and inserted into pOB150. The GEST coding sequence was thus fused to GFP, the plasmid also contained the human dhfr resistance marker. | ||||||||||||||||||||||||||
| Additional remarks selection procedure | |||||||||||||||||||||||||||
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