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Details of the target gene |
Gene Model of Rodent Parasite |
PBANKA_0522700
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Gene Model P. falciparum ortholog |
PF3D7_0422300
|
Gene product | alpha tubulin 2 |
Gene product: Alternative name | |
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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 used | (Linear) 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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Partial or complete disruption of the gene | Complete |
Additional remarks partial/complete disruption |
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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 | Alpha-tubulin II is one of the two alpha-tubulin genes (I and II) in Plasmodium. This gene is highly expressed in male gametocytes. Expression was also observed in the asexual blood stages, female gametocytes, ookinetes and oocysts (additional P. berghei gene models: PB001519.02.0-PB300720.00.0-PB000609.00.0). The paper describes two different constructs that have been used in attempts to disrupt alpha-tubulin ii; pL0103 (for details see below) and pL1006 (details not shown). |
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 | CCCAAGCTTGGATCCTGTAGATATATCCACATTTTACA |
Additional information primer 1 | Primer L468; 5'UTR Pb alpha-tub II |
Sequence Primer 2 | CCCAAGCTTGCTAATAACTTCTCTCATTTTCG |
Additional information primer 2 | Primer L469; 5'UTR Pb alpha-tub II |
Sequence Primer 3 | GGATATCCATCACCACAGGTTTCTACTGC |
Additional information primer 3 | Primer L470; Exon 3 Pb alpha-tub I & II |
Sequence Primer 4 | GGAATTCAAACTTCAGCAATAGCAGTTGAG |
Additional information primer 4 | Primer L471; Exon 3 Pb alpha-tub I & II |
Sequence Primer 5 | |
Additional information primer 5 | |
Sequence Primer 6 | |
Additional information primer 6 | |
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