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Type and details of transgene |
Is the transgene Plasmodium derived |
Transgene: not Plasmodium |
Transgene name | ORP1 (PBANKA_0902500) fused to GFP |
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Details of the genetic modification |
Inducable system used | No |
Additional remarks inducable system |
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Type of plasmid/construct | (Linear) plasmid single 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 plasmid used for the generation ORP1::GFP transgenic parasites is a derivative from the pDEF-PbDHFR-SSU (targeting the C/D ssu ribosomal RNA locus by single cross-over integration).
ORP1::GFP is expressed as an extra copy of the gene in the C/D ssu ribosomal RNA locus. The promoter region of 1,232bp was amplified using primers TF50for and TF50rev, and inserted in the sites XhoI–XmaI. The fragment corresponding to the first 365 aa of the coding region were amplified by primers TF260for and TF260rev, and inserted using restriction enzymes XmaI–ApaI. The 482bp ORP1 30-FR region was amplified by TF30 for and TF30rev and inserted in the NheI and NotI restriction sites. |
Additional remarks selection procedure | |
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Other details transgene |
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Promoter |
Gene Model of Parasite |
PBANKA_0902500
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Gene Model P. falciparum ortholog |
PF3D7_1146600
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Gene product | oocyst rupture protein 1, putative | CCAAT-box DNA binding protein subunit B |
Gene product: Alternative name | ORP1; NFYB |
Primer information details of the primers used for amplification of the promoter sequence
Primer information details of the primers used for amplification of the promoter sequence
Sequence Primer 1 | |
Additional information primer 1 | |
Sequence Primer 2 | |
Additional information primer 2 | |
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3'-UTR |
Gene Model of Parasite |
PBANKA_0902500
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Gene product | oocyst rupture protein 1, putative; CCAAT-box DNA binding protein subunit B |
Gene product: Alternative name | ORP1; NFYB |
Primer information details of the primers used for amplification the 3'-UTR sequences
Primer information details of the primers used for amplification the 3'-UTR sequences
Sequence Primer 1 | |
Additional information primer 1 | |
Sequence Primer 2 | |
Additional information primer 2 | |
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Insertion/Replacement locus |
Replacement / Insertion | Insertion locus |
Gene Model of Parasite |
Not available
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Gene product | Not available |
Gene product: Alternative name | small subunit ribosomal rna gene (c/d-type unit) |
Primer information details of the primers used for amplification of the target sequences
Primer information details of the 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 | |
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