SummaryRMgm-352
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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 | P. berghei ANKA cl15cy1 | ||||||||||||||||||||||||
Other information parent line | A reference wild type clone from the ANKA strain of P. berghei (PubMed: PMID: 17406255). | ||||||||||||||||||||||||
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The mutant parasite was generated by | |||||||||||||||||||||||||
Name PI/Researcher | B. Franke-Fayard; C.J. Janse | ||||||||||||||||||||||||
Name Group/Department | Leiden Malaria Research Group | ||||||||||||||||||||||||
Name Institute | Leiden University Medical Center | ||||||||||||||||||||||||
City | Leiden | ||||||||||||||||||||||||
Country | The Netherlands | ||||||||||||||||||||||||
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Name of the mutant parasite | |||||||||||||||||||||||||
RMgm number | RMgm-352 | ||||||||||||||||||||||||
Principal name | 314cl1 | ||||||||||||||||||||||||
Alternative name | Δp230p-II | ||||||||||||||||||||||||
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 | Not different from wild type | ||||||||||||||||||||||||
Liver stage | Not different from wild type | ||||||||||||||||||||||||
Additional remarks phenotype | Mutant/mutation The p230p locus is commonly used as a locus for integration of transgenes in P. berghei, for example fluorescent proteins. The mutant described here, Δp230p-II, contains a partial disrupted p230p gene. A fragment of the gene is deleted that includes part of the first 6-cys domain (i.e. the first 492 amino acids are still present in the genome). A second mutant (Δp230p-I; RMgm-351) has been generated and a fragment of the gene is deleted from the second 6-cys domain (i.e. the first 894 amino acids are still present. Table: P. falciparum gene members of the 6-cys family
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Details of the target gene | |||||||||||||||||||||||||
Gene Model of Rodent Parasite | PBANKA_0306000 | ||||||||||||||||||||||||
Gene Model P. falciparum ortholog | PF3D7_0208900 | ||||||||||||||||||||||||
Gene product | 6-cysteine protein | ||||||||||||||||||||||||
Gene product: Alternative name | P230p | ||||||||||||||||||||||||
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Details of the genetic modification | |||||||||||||||||||||||||
Inducable system used | No | ||||||||||||||||||||||||
Additional remarks inducable system | |||||||||||||||||||||||||
Type of plasmid/construct used | Plasmid double cross-over | ||||||||||||||||||||||||
PlasmoGEM (Sanger) construct/vector used | No | ||||||||||||||||||||||||
Modified PlasmoGEM construct/vector used | No | ||||||||||||||||||||||||
Plasmid/construct map | |||||||||||||||||||||||||
Plasmid/construct sequence |
AATTCACTGGCCGTCGTTTTACAACGTCGTGACTGGGAAAACCCTGGCGTTACCCAACTT
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Restriction sites to linearize plasmid | SacII | ||||||||||||||||||||||||
Partial or complete disruption of the gene | Partial | ||||||||||||||||||||||||
Additional remarks partial/complete disruption | Of the 6863 coding region, nucleotides 2166-3317 were disrupted. | ||||||||||||||||||||||||
Selectable marker used to select the mutant parasite | tgdhfr | ||||||||||||||||||||||||
Promoter of the selectable marker | pbdhfr | ||||||||||||||||||||||||
Selection (positive) procedure | pyrimethamine | ||||||||||||||||||||||||
Selection (negative) procedure | No | ||||||||||||||||||||||||
Additional remarks genetic modification | The 230p locus was disrupted by introduction of the Tgdhfr selectable marker and GFPmut3, both under control of the eef1a promoter. GFP is expressed in all life cycle stages except for male gametes. | ||||||||||||||||||||||||
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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