RMgmDB - Rodent Malaria genetically modified Parasites
Back to search resultsSummaryRMgm-4144
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Last modified: 18 May 2017, 12:20
*RMgm-4144| Successful modification | The gene/parasite could not be changed/generated by the genetic modification. |
| The following genetic modifications were attempted | Gene disruption |
| Number of attempts to introduce the genetic modification | 3 |
| Reference (PubMed-PMID number) |
Reference 1 (PMID number) : 28481199 |
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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 820cl1m1cl1 (RMgm-164) |
| Other information parent line | P. berghei ANKA 820cl1m1cl1 (RMgm-164) is a reference ANKA mutant line which expresses GFP under control of a male and RFP under control of a female gametocyte specific promoter. This reference line does not contain a drug-selectable marker (PubMed: PMID: 19438517). |
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| Attempts to generate the mutant parasite were performed by | |
| Name PI/Researcher | Fang H, Brochet M |
| Name Group/Department | Department of Microbiology and Molecular Medicine |
| Name Institute | University of Geneva |
| City | Geneva |
| Country | Switzerland |
Disrupted: Mutant parasite with a disrupted gene| top of page | |||||||||||||||||||||||||
| Details of the target gene | |||||||||||||||||||||||||
| Gene Model of Rodent Parasite | PBANKA_1115300 | ||||||||||||||||||||||||
| Gene Model P. falciparum ortholog | PF3D7_0515700 | ||||||||||||||||||||||||
| Gene product | glideosome-associated protein 40, putative | ||||||||||||||||||||||||
| Gene product: Alternative name | GAP40 | ||||||||||||||||||||||||
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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 | Yes | ||||||||||||||||||||||||
| Name of PlasmoGEM construct/vector | PbGEM-335107 | ||||||||||||||||||||||||
| 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 protein is identified in a screen for proteins that are phosphorylated by CDPK4.CDPK4 has an essential role in male (micro)gamete formation (exflagellation). Among these proteins, it was decided to name those without predicted function SOC, for substrate of CDPK4. Ten proteins were indeed conserved Plasmodium proteins of unknown function and one protein, SOC2, was annotated as cyclin-related protein 2 but it was suggested be unrelated to cyclins. Also GAP40 was identified. To investigate the roles of SOC1 to 4 and GAP40 during gametogenesis, it was attempted to individually knock-out their encoding proteins. As controls, soc5 and soc6 were included that were not differentially phosphorylated in a CDPK4-KO mutant. For gap40 no transgenic parasites could be obtained. Viable KO parasites could be detected in mixed asexual stages for six genes and non-clonal populations were first assessed for microgametocyte DNA replication and exflagellation. No defects were observed in the SOC4-KO line as well as in the SOC5-KO and SOC6-KO control lines suggesting that these proteins do not represent crucial CDPK4 effectors in the regulation of male gametogenesis. Defects in DNA replication or exflagellation were observed for SOC1-KO, SOC2-KO, and SOC3-KO lines that were cloned for in depth characterisation. | ||||||||||||||||||||||||
| Additional remarks selection procedure | |||||||||||||||||||||||||
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Primer information: Primers used for amplification of the target sequences
 Primer information: Primers used for amplification of the target sequences
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