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Details of the target gene |
Gene Model of Rodent Parasite |
PBANKA_1459500
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Gene Model P. falciparum ortholog |
PF3D7_1246400
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Gene product | myosin light chain 1 | myosin A tail domain interacting protein |
Gene product: Alternative name | MLC1; MTIP |
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Details of the genetic modification |
Short description of the mutation | 'Promoter swap' mutant: the promoter of MTIP replaced by the promoter of ama1 (PBANKA_091500) |
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 | hdhfr |
Promoter of the selectable marker | pbdhfr |
Selection (positive) procedure | pyrimethamine |
Selection (negative) procedure | No |
Additional remarks genetic modification | For the double-crossover promoter-exchange plasmid, a parent vector Pama1 was first constructed containing the ama1 promoter. For Pama1, 1.5 kb of the upstream sequence of the ama1 gene (PBANKA_083630) was cloned downstream of the hdhfr cassette in pDEFhDHPEA. Pama1-mtip (pSS369) vector was subsequently made by inserting a mtip 5’ homology region upstream of the hdhfr cassette in Pama1 (consisting of 500 bp of mtip upstream sequence), and a 3’ homology region downstream of the ama1 promoter (consisting of the first 500 bp of mtip coding sequence). |
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 | gagaccgcggttactatatatgtgtagaaatg |
Additional information primer 1 | olSS1016; 5’HR MTIP forward |
Sequence Primer 2 | gagactgcagttttgcgatatatattttttaaattaa |
Additional information primer 2 | olSS1017; 5’HR MTIP reverse |
Sequence Primer 3 | gagactcgagatggaacaacaatgccatat |
Additional information primer 3 | olSS1018; 3’HR MTIP forward |
Sequence Primer 4 | gagagcggccgcgatatcctcgcaaaataatttataat |
Additional information primer 4 | olSS1019; 3’HR MTIP reverse |
Sequence Primer 5 | |
Additional information primer 5 | |
Sequence Primer 6 | |
Additional information primer 6 | |
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