SummaryRMgm-5429
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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) | Not published (yet) |
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 | Hentzschel F, Frischknecht F |
Name Group/Department | Heidelberg University Medical Faculty, University Hospital Heidelberg |
Name Institute | Center for Infectious Diseases |
City | Heidelberg |
Country | Germany |
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Name of the mutant parasite | |
RMgm number | RMgm-5429 |
Principal name | spm1(-) |
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 different from wild type |
Sporozoite | Not different from wild type |
Liver stage | Not different from wild type |
Additional remarks phenotype | Mutant/mutation |
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Details of the target gene | |||||||||||||||||||||||||
Gene Model of Rodent Parasite | PBANKA_0810700 | ||||||||||||||||||||||||
Gene Model P. falciparum ortholog | PF3D7_0909500 | ||||||||||||||||||||||||
Gene product | subpellicular microtubule protein 1, putative | ||||||||||||||||||||||||
Gene product: Alternative name | SPM1 | ||||||||||||||||||||||||
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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 | - | ||||||||||||||||||||||||
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 | For gene deletion of PbSPM1 (PBANKA_0810700), PbTrxL1 (PBANKA_0820200), PbFAP20 (PBANKA_1108000) and PbFAP52 (PBANKA_1361500), the respective transfection vectors were obtained from PlasmoGEM. Prior to transfection, 3-5 µg of each vector were linearized using NotI followed by ethanol precipitation. The linearized vectors were transfected into the parental P. berghei strain (PbANKA wildtype for SPM1 and TrxL1, and Pb820 wildtype for FAP20 and FAP50). In order to be able to reuse pyrimethamine as a selective (positive) pressure in subsequent mutant generations, 5-fluorocytosine (5-FC) selection (negative) was performed. Here, trxL1(-) and fap20(-) parasites that recycled the selection cassette were negatively selected for via 5-FC (1 mg/ml) administered via the mice’s drinking water and isogenic clones were isolated by limited dilution and verified via genotyping PCR accordingly. After obtaining marker-free clones, the second knockout (spm1 or fap52) was generated in these lines. | ||||||||||||||||||||||||
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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