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Details of the target gene |
Gene Model of Rodent Parasite |
PBANKA_0403200
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Gene Model P. falciparum ortholog |
PF3D7_0304600
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Gene product | circumsporozoite (CS) protein |
Gene product: Alternative name | CS; CSP |
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Details of the genetic modification |
Short description of the mutation | Chimeric cs gene: the repeat region of cs is replaced with the repeat region of P. vivax (VK210) cs |
Inducable system used | No |
Short description of the conditional mutagenesis | Not available |
Additional remarks inducable system |
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Type of plasmid/construct | Plasmid double cross-over |
PlasmoGEM (Sanger) construct/vector used | No |
Modified PlasmoGEM construct/vector used | No
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Plasmid/construct map |
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Plasmid/construct sequence |
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Restriction sites to linearize plasmid |
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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 | The transfection plasmid, pR-CSPv, carrying the repeat region of P. vivax was derived from plasmid pR-CSwt that encodes for the CS gene of P. berghei (wildtype) and results from the combination of plasmids pIC-CSwt and a synthetic gene pPv (Retrogen Inc., CA) encoding the VK210 repeat motif based on the Salvador I (Sal I) strain of P. vivax. Briefly, a PmlI-SexAI 567-bp fragment was excised from pIC-CSwt and replaced with a PmlI-SexAI 675-bp fragment, comprising the P. vivax (VK210) CSP repeat region that was codon harmonized to optimize protein synthesis and folding in the P. berghei transgenic host and was excised from the engineered pPv. From the resulting intermediate plasmid, pIC-CSPv, the entire CSP gene was excised as a KpnI-XhoI fragment and inserted into the transfection plasmid, pR-CSPv. KpnI and SacI were used to linearize pR-CSPv prior to transfection. |
Additional remarks selection procedure | |
Primer information: Primers used for amplification of the target sequences
Primer information: Primers used for amplification of the target sequences
Sequence Primer 1 | |
Additional information primer 1 | |
Sequence Primer 2 | |
Additional information primer 2 | |
Sequence Primer 3 | |
Additional information primer 3 | |
Sequence Primer 4 | |
Additional information primer 4 | |
Sequence Primer 5 | |
Additional information primer 5 | |
Sequence Primer 6 | |
Additional information primer 6 | |
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