top of page |
Details of the target gene |
Gene Model of Rodent Parasite |
PBANKA_1133300
|
Gene Model P. falciparum ortholog |
PF3D7_1357100
|
Gene product | elongation factor 1-alpha |
Gene product: Alternative name | eef1ab, elongation factor 1 alpha b |
top of page |
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 |
|
Restriction sites to linearize plasmid |
BamHI/EcoRI
|
Partial or complete disruption of the gene | Partial |
Additional remarks partial/complete disruption |
eef1ab was disrupted by inserting the Tgdhfr selectable marker into the ORF replacing 151 bp of its central coding region. eef1a promoter and eef1aa remained intact.
|
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 | This mutant exhibited an unpredicted integration event. The eef1ab gene is disrupted, but these parasites contain an intergenic region that is inverted compared with the arrangement of the pbeef1a intergenic region of wt parasites (see also mutant 174 which also contains a disrupted eef1ab gene but with a normal organization of the intergenic region). |
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 | CCCAAGCTTGGATCCACTAGTATGGGAAAAGAAAAAACTCAC |
Additional information primer 1 | L402 (HindIII,BamHI,SpeI); Pbef-1α 5' end ORF |
Sequence Primer 2 | CCCAAGCTTACTAGTCGGCTTGATATCCTACC |
Additional information primer 2 | L403 (HIII, SpeI); Pbef-1α 5' end ORF |
Sequence Primer 3 | GGATATCTTCCATTACAAGGTGTAT |
Additional information primer 3 | L404 (EcoRV); Pbef-1α 3' end ORF |
Sequence Primer 4 | GGGGTACCGCTGGTGCTTTAGCTGAG |
Additional information primer 4 | L405 (KpnI); Pbef-1α 3' end ORF |
Sequence Primer 5 | |
Additional information primer 5 | |
Sequence Primer 6 | |
Additional information primer 6 | |
|
top of page |