Summary

RMgm-513
Malaria parasiteP. berghei
Genotype
DisruptedGene model (rodent): PBANKA_1233500; Gene model (P.falciparum): PF3D7_0518700; Gene product: mRNA-binding protein PUF1 (Pumilio1)
Phenotype Fertilization and ookinete;
Last modified: 1 June 2011, 16:31
  *RMgm-513
Successful modificationThe parasite was generated by the genetic modification
The mutant contains the following genetic modification(s) Gene disruption
Reference (PubMed-PMID number) Reference 1 (PMID number) : 21625527
MR4 number
Parent parasite used to introduce the genetic modification
Rodent Malaria ParasiteP. berghei
Parent strain/lineP. berghei ANKA
Name parent line/clone P. berghei ANKA cl15cy1
Other information parent lineA reference wild type clone from the ANKA strain of P. berghei (PubMed: PMID: 17406255).
The mutant parasite was generated by
Name PI/ResearcherC.S.S Gomes-Santos; J.A.M. Braks: G.R. Mair
Name Group/DepartmentMalaria Unit
Name InstituteInstituto de Medicina Molecular
CityLisbon
CountryPortugal
Name of the mutant parasite
RMgm numberRMgm-513
Principal name351cl1
Alternative namePbpuf1-KOa
Standardized name
Is the mutant parasite cloned after genetic modificationYes
Phenotype
Asexual blood stageNot different from wild type
Gametocyte/GameteNot different from wild type
Fertilization and ookineteNormal numbers of gametocytes and ookinetes are produced (see also 'Additional remarks phenotype').
OocystNot different from wild type
SporozoiteNot different from wild type
Liver stageNot different from wild type
Additional remarks phenotype

Mutant/mutation
The mutant lacks expression of the RNA binding protein Puf1.

Protein (function)
The roles of Puf (Pumilio and fem-3 mRNA binding factor) proteins are diverse yet intimately involved in the translational regulation of developmental and differentiation factors in organisms as diverse as yeast, C. elegans, Drosophila and humans. Two such proteins, Puf1 (PFE0935c) and Puf2 (PFD0825c), are known in the human malaria parasite, P. falciparum, with orthologs in all Plasmodium spp. characterized. The Plasmodium Puf proteins have the typical highly conserved organization that includes the eight tandem copies of the PUM RNA binding domain (or Pumilio Homology Domain, PHD) at the carboxyterminus of the protein and expectedly PfPuf2 was shown to bind RNA in vitro. In P. falciparum evidence has been reported for a role for Puf2 in gametocyte development although Pfpuf2 is most highly transcribed in sporozoites (see also 'Additional Information').

Phenotype
The phenotype analyses indicate that Puf1 is not essential for blood stage development, production of gametocytes, ookinetes, oocysts and infective sporozoites and for mosquito transmission and liver stage development (see also 'Additional information').

Additional information
Normal numbers of gametocytes and ookinetes are produced. Mutant ookinetes showed a slightly different morphology at the light-microscope level (more 'stumpy and rounder' forms compared to the banana-shaped wild type ookinetes). The morphology of mutant ookinetes have not been analysed in more detail.

See mutant RMgm-515, RMgm-516 and RMgm-591 for P. berghei mutants lacking expression PUF2. These mutants show normal blood stage development, gametocyte, ookinete and oocyst production but sporozoite infectivity is affected. 

Mutant RMgm-591 that lacks expression of both Puf1 and Puf2 shows the same phenotype as mutants lacking only expression of Puf2.

In P. falciparum Puf2 expression has been demonstrated in gametocytes and disruption of Pfpuf2 (PFD0825c) resulted in increased gametocyte production and increased differentiation of male gametocytes (Miao J. et al., 2010, 123-1039-1049). The phenotype of the P. falciparum mutants lacking expression of Puf2 has not been analysed during mosquito development.

Other mutants
RMgm-514: An independent mutant lacking expression of Puf1 which expresses GFP under the constitutive eef1α promoter
RMgm-515: A mutant lacking expression of Puf2 (PBANKA_071920)
RMgm-516: An independent mutant lacking expression of Puf2 (PBANKA_071920) which expresses GFP under the constitutive eef1α promoter
RMgm-591: A mutant lacking expression of both Puf1 and Puf2 which expresses GFP under the constitutive eef1α promoter


  Disrupted: Mutant parasite with a disrupted gene
Details of the target gene
Gene Model of Rodent Parasite PBANKA_1233500
Gene Model P. falciparum ortholog PF3D7_0518700
Gene productmRNA-binding protein PUF1
Gene product: Alternative namePumilio1
Details of the genetic modification
Inducable system usedNo
Additional remarks inducable system
Type of plasmid/construct usedPlasmid double cross-over
PlasmoGEM (Sanger) construct/vector usedNo
Modified PlasmoGEM construct/vector usedNo
Plasmid/construct map
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Plasmid/construct sequence
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GTCAACAATAAATAATAAATAAATATTGTGGAAATAAAATAACATATAATTATTTTTAAT
ACATTGATTTCCCTTTTATTTTTTTAAATTTCATTGATATAAAAATATATAATAATAACA
TATATGATTTCAAATTAATCTTTTCAAAAATGGTGTTTATTTTTGTATGTTTGTGTATGA
ATTAATCACATAACACATCTATTAAATTGAGTTGGTAATATAGACACAAATAAATATATA
TATTTTTATAGCTTAAAAGTGTGTTATGAATATTTTAAGCATATTTTCTTTTTCTTTGGA
TTGTGTAAAATGAACTCATATAATGCGTTTTTTTGTTTTTGTTATTTTGTCATTTTGTTA
TTTTGCTATTTTATGGATTAATTTTTGTTTATAAAATGGGAAATAATTTTAACATATTTA
AATAAATGGAGAAAAAATATAAAATAATTATAAAAAAAAGTTAATACACATTTTTTCCTG
TTATAGACCTTATATTTATTTATCCATATATATATATATATATATATATATATATATATA
CATACCAAGTGAATTAAGAGGAAAGCTAATTTATTATTCAGAATAATATATGAACTATAT
ATAATTTTTATTATTTTGGTGTATATTAATCTGTCTATATGCATACATGCAATAATTTAT
CGACTTATATATCAAATAACATAAAATAGAAGTGTTTTAAATTATGGATATATGCTCAAT
ATTCATTTTTTTTAATAAGTTAGCTATATTTAAATTATACATTTTATATATGGTCTCTTT
TTTTTTTAAATATTATTTAAGTGATCATGAAAATATAAATAATTTTTTTTTATTTAATAT
CCTTTTGCTTGCATGTGGTAAATGGAAATTTGGATGTGTTTTGAARGTTCGGATATAGTT
GTATGGACATATAATATATTTTGTGAAAAATTGGTTTTATGTTTATACTTATGCCAATAC
TTTTTGAGTAAAACAAAGCAAGTGCTTATAAATAATTAAAGCCAATTTTATAATATATAT
TTTTTTATTTAATTTGAATTTAGTAGTATAATTTTTTATGGTAAGTGCTCAAAGAGAGTT
GCTTATAAAGTATGGTTTGTTTCTTTTTCGCCATTTTGAATTACACATTAAAAATATATA
GATACATATATTATAATATGAAATCATTAATAATTTAGGGAAATTCTACAAATTTAAAAA
CGAATAAAATAATTGTTTTTCATCATGCCATAACACAATATTGATATATACATGTACAAA
CATTTTTTTTATTTGGAAAATATAAATTATATAAAAAAAAATGTATAGTATACAAAATGA
GCATATTCACACGGGGTGGACGTTCATTTTTTCATTTTTCCCCTGTTTTTTATGAGTATA
TGATAAAATTTTATGAACATTTACACAAAATGAAAATGGATATATAGGAAAAATGGAGCG
GTATTTCATTTATCTTTGATTGTCATTTGGATATTATATTACCYTGGGTAGGCAATTAAA
AATGTTAAATAACAATTTAAGGAAATTATATTTTATATATTAAAATTAACACTGTATTAT
ATGATTCGCTTATAAAAGCCACTCTTTCCCCATGCAAAGCTGTTTAATATCAATTTTAAC
AAATTACACACATGTTAATATATTTATATATATAATTTATATATTTATAATTTATATATT
TATATTTTTATTATTTATATATTTATTATTTATTGTGTGTGTCAATTCGGGTAGGATATA
CCTCTTTTTTATTGTTTAAAGCGATTTGTATTCTAAAATATAAAGRATTTGAAAAAGAGA
AAGATAGAATATGATCCCATCATATATAGCCCTATAATTTTTATTTAGCAGCGAATTAAT
TTTTCTATTAAGTTTATGTGTAATTAAAATAACGGAATATATATAATACAATAAAAAAGT
GCATAAATTAAAATTTTTTCAATTAAATTTTTTTTTTTAAGGGGTTATATAATATTAAAT
ATATAAAATACGATTATATATTTTTGCTACAATTTTTTATATTAAGATATAAATAGTAAA
TAAATGGTATTATATGGCATGTAATATATAAATTTTTTCCAATTTTTATTTTATATACAC
TTTTCCTTTTTTTGTCATAAAACTTAAACAATTTACACATTCATTTTAAAAATTGACTAT
TTGTTTCAACATTTTTTGAGTTTCCGTTTTATAATAGTATTTTCATTTGTATATTGCTTA
TATATATAAATACACACCTAAATGTTACAAAGGATCAATGCATAAACCGGTGTGTCTGGT
CGTCGCGATGACCCCCAAGAGGGGCATCGGCATCAACAACGGCCTCCCGTGGCCCCACTT
GACCACAGATTTCAAACACTTTTCTCGTGTGACAAAAACGACGCCCGAAGAAGCCAGTCG
CCTGAACGGGTGGCTTCCCAGGAAATTTGCAAAGACGGGCGACTCTGGACTTCCCTCTCC
ATCAGTCGGCAAGAGATTCAACGCCGTTGTCATGGGACGGAAAACCTGGGAAAGCATGCC
TCGAAAGTTTAGACCCCTCGTGGACAGATTGAACATCGTCGTTTCCTCTTCCCTCAAAGA
AGAAGACATTGCGGCGGAGAAGCCTCAAGCTGAAGGCCAGCAGCGCGTCCGAGTCTGTGC
TTCACTCCCAGCAGCTCTCAGCCTTCTGGAGGAAGAGTACAAGGATTCTGTCGACCAGAT
TTTTGTCGTGGGAGGAGCGGGACTGTACGAGGCAGCGCTGTCTCTGGGCGTTGCCTCTCA
CCTGTACATCACGCGTGTAGCCCGCGAGTTTCCGTGCGACGTTTTCTTCCCTGCGTTCCC
CGGAGATGACATTCTTTCAAACAAATCAACTGCTGCGCAGGCTGCAGCTCCTGCCGAGTC
TGTGTTCGTTCCCTTTTGTCCGGAGCTCGGAAGAGAGAAGGACAATGAAGCGACGTATCG
ACCCATCTTCATTTCCAAGACCTTCTCAGACAACGGGGTTCCCTACGACTTTGTGGTTCT
CGAGAAGAGAAGGAAGACTGACGACGCAGCCACTGCGGAACCGAGCAACGCAATGAGCTC
CTTGACGTCCACGAGGGAGACAACTCCCGTGCACGGGTTGCAGGCTCCTTCTTCGGCCGC
AGCCATTGCCCCGGTGTTGGCGTGGATGGACGAAGAAGACCGGAAAAAACGCGAGCAAAA
GGAACTGATTCGGGCCGTTCCGCATGTTCACTTTAGAGGCCATGAAGAGTTCCAGTACCT
TGATCTCATTGCCGACATTATTAACAATGGAAGGACAATGGATGACCGAACGGGCGTTGG
TGTCATCTCCAAATTCGGCTGCACTATGCGCTACTCGCTGGATCAGGCCTTTCCACTTCT
CACCACAAAGCGTGTGTTCTGGAAAGGGGTCCTCGAAGAGTTGCTGTGGTTCATTCGCGG
CGACACGAACGCAAACCATCTTTCTGAGAAGGGCGTGAAGATCTGGGACAAGAATGTGAC
ACGCGAGTTCCTCGATTCGCGCAATCTCCCCCACCGAGAGGTCGGAGACATCGGCCCGGG
CTACGGCTTCCAGTGGAGACACTTCGGCGCGGCATACAAAGACATGCACACAGACTACAC
AGGGCAGGGCGTCGACCAGCTGAAGAATGTGATCCAGATGCTGAGAACGAATCCAACAGA
TCGTCGCATGCTCATGACTGCCTGGAATCCTGCAGCGCTGGACGAAATGGCGCTGCCGCC
TTGTCACTTGTTGTGCCAGTTCTACGTGAACGACCAGAAGGAGCTGTCGTGCATCATGTA
TCAGCGGTCGTGCGATGTCGGCCTCGGCGTCCCCTTCAACATCGCTTCCTATTCGCTTTT
GACGCTCATGGTTGCACACGTCTGCAACCTAAAACCTAAGGAGTTCATTCACTTCATGGG
GAACACGCATGTCTACACGAACCATGTCGAGGCTTTAAAAGAGCAGCTGCGGAGAGAACC
GAGACCGTTCCCCATTGTGAACATCCTCAACAAGGAACGCATCAAGGAAATCGACGATTT
CACCGCCGAGGATTTTGAGGTCGTGGGCTACGTCCCGCACGGACGAATCCAGATGGAGAT
GGCTGTCTAGCGGAAATACAGAAGCTGGCTTTGATCCCGTTTTTCTTACTTATATATTTA
TACCAATTGATTGTATTTATAACTGTAAAAATGTGTATGTTGTGTGCATATTTTTTTTTG
TGCATGCACATGCATGTAAATAGCTAAAATTATGAACATTTTATTTTTTGTTCAGAAAAA
AAAAACTTTACACACATAAAATGGCTAGTATGAATAGCCATATTTTATATAAATTAAATC
CTATGAATTTATGACCATATTAAAAATTTAGATATTTATGGAACATAATATGTTTGAAAC
AATAAGACAAAATTATTATTATTATTATTATTTTTACTGTTATAATTATGTTGTCTCTTC
AATGATTCATAAATAGTTGGACTTGATTTTTAAAATGTTTATAATATGATTAGCATAGTT
AAATAAAAAAAGTTGAAAAATTAAAAAAAAACATATAAACACAAATGATGTTTTTTCCTT
CAATTTCGATATCGTTCCTTGCATGTGTGTGTAGAATTATTATCATTTATTCTATCAGCG
GAATCAAAATAATTGGATAAATTATTCTTACTATAATTTATCGTACTCATTGCGTCCATT
GTATTTACACCATTTGGTGGGCAATGTACTATTGGTGTCATATTATTATTTATAAAATCT
TTATTTAATATATATTTATTATTATATTTTTTATATGAACTGCTCATGTAATTTTGTAGA
TTGTAATTCATGCTATGTGTATTCACATATGGAATATTTATTTGATATAGTTGTGTATAA
TTTGATATGTTGGGTAATGCTACTCTATAATTCGGTGGTATTATATCTATTGGATTCATA
ATTTTTGTATTTATATTTGAATCAGGTATTTTATTTATCGGTGTATTTTTAGTTTTTTTT
TTTTTATTACTTATTTCATCTGTCTTCGCTAAAGAATCATCATCAGGTATTTTCTTATTT
TTATTCATTGAATTTGAAGTGTCTTTATTATTGCATTTATCTTCACTGCTATTACAACTC
TCGGTATTGCTCTCAAAGTTACTACTCTTAAAGATTTCCTTATTCACATCATGTATATTT
TTCATTTTTTCGGTATTTTCGAAATTTATTGTATCATTAATATCTTTGTCAGTTTTATTC
TCTAGTTCATTTTCACAAACTAAATTATCTGATGACAATTCTGGTGATGTTTCTCGCTTC
TCATTTTTATTTTCCATTTTATCCGAACTTATTCGTTTCTTTATTTCTTTCGTTTTTTTT
CCTTTTTTGGTTTTTGGTGAATAGGGAATGGGAAGGTATATTTTTATATTCGGATTAATT
AAAAAAATAAAAGCAAAAAATAGTTTAGTTACATAAAAAAAATTGTTGCAAAAACATTTT
TATAAATGATAAAGATAAAAAAACATATTGGATTGTATAATTATTTATTGTCTAAAAATT
TAAATACACATAAAAAACTCAAACACTTCTATATATGTGCGTAATGCTAAGCATTGATAT
TTACGAATTTTCAATGTGTTTTGTTCGGATCCACTAGTTCTAGAGCGGCCGCCACCGCGG
TGGAGCTCCAGCTTTTGTTCCCTTTAGTGAGGGTTAATTTCGAGCTTGGCGTAATCATGG
TCATAGCTGTTTCCTGTGTGAAATTGTTATCCGCTCACAATTCCACACAACATACGAGCC
GGAAGCATAAAGTGTAAAGCCTGGGGTGCCTAATGAGTGAGCTAACTCACATTAATTGCG
TTGCGCTCACTGCCCGCTTTCCAGTCGGGAAACCTGTCGTGCCAGCTGCATTAATGAATC
GGCCAACGCGCGGGGAGAGGCGGTTTGCGTATTGGGCGCTCTTCCGCTTCCTCGCTCACT
GACTCGCTGCGCTCGGTCGTTCGGCTGCGGCGAGCGGTATCAGCTCACTCAAAGGCGGTA
ATACGGTTATCCACAGAATCAGGGGATAACGCAGGAAAGAACATGTGAGCAAAAGGCCAG
CAAAAGGCCAGGAACCGTAAAAAGGCCGCGTTGCTGGCGTTTTTCCATAGGCTCCGCCCC
CCTGACGAGCATCACAAAAATCGACGCTCAAGTCAGAGGTGGCGAAACCCGACAGGACTA
TAAAGATACCAGGCGTTTCCCCCTGGAAGCTCCCTCGTGCGCTCTCCTGTTCCGACCCTG
CCGCTTACCGGATACCTGTCCGCCTTTCTCCCTTCGGGAAGCGTGGCGCTTTCTCATAGC
TCACGCTGTAGGTATCTCAGTTCGGTGTAGGTCGTTCGCTCCAAGCTGGGCTGTGTGCAC
GAACCCCCCGTTCAGCCCGACCGCTGCGCCTTATCCGGTAACTATCGTCTTGAGTCCAAC
CCGGTAAGACACGACTTATCGCCACTGGCAGCAGCCACTGGTAACAGGATTAGCAGAGCG
AGGTATGTAGGCGGTGCTACAGAGTTCTTGAAGTGGTGGCCTAACTACGGCTACACTAGA
AGGACAGTATTTGGTATCTGCGCTCTGCTGAAGCCAGTTACCTTCGGAAAAAGAGTTGGT
AGCTCTTGATCCGGCAAACAAACCACCGCTGGTAGCGGTGGTTTTTTTGTTTGCAAGCAG
CAGATTACGCGCAGAAAAAAAGGATCTCAAGAAGATCCTTTGATCTTTTCTACGGGGTCT
GACGCTCAGTGGAACGAAAACTCACGTTAAGGGATTTTGGTCATGAGATTATCAAAAAGG
ATCTTCACCTAGATCCTTTTAAATTAAAAATGAAGTTTTAAATCAATCTAAAGTATATAT
GAGTAAACTTGGTCTGACAGTTACCAATGCTTAATCAGTGAGGCACCTATCTCAGCGATC
TGTCTATTTCGTTCATCCATAGTTGCCTGACTCCCCGTCGTGTAGATAACTACGATACGG
GAGGGCTTACCATCTGGCCCCAGTGCTGCAATGATACCGCGAGACCCACGCTCACCGGCT
CCAGATTTATCAGCAATAAACCAGCCAGCCGGAAGGGCCGAGCGCAGAAGTGGTCCTGCA
ACTTTATCCGCCTCCATCCAGTCTATTAATTGTTGCCGGGAAGCTAGAGTAAGTAGTTCG
CCAGTTAATAGTTTGCGCAACGTTGTTGCCATTGCTACAGGCATCGTGGTGTCACGCTCG
TCGTTTGGTATGGCTTCATTCAGCTCCGGTTCCCAACGATCAAGGCGAGTTACATGATCC
CCCATGTTGTGCAAAAAAGCGGTTAGCTCCTTCGGTCCTCCGATCGTTGTCAGAAGTAAG
TTGGCCGCAGTGTTATCACTCATGGTTATGGCAGCACTGCATAATTCTCTTACTGTCATG
CCATCCGTAAGATGCTTTTCTGTGACTGGTGAGTACTCAACCAAGTCATTCTGAGAATAG
TGTATGCGGCGACCGAGTTGCTCTTGCCCGGCGTCAATACGGGATAATACCGCGCCACAT
AGCAGAACTTTAAAAGTGCTCATCATTGGAAAACGTTCTTCGGGGCGAAAACTCTCAAGG
ATCTTACCGCTGTTGAGATCCAGTTCGATGTAACCCACTCGTGCACCCAACTGATCTTCA
GCATCTTTTACTTTCACCAGCGTTTCTGGGTGAGCAAAAACAGGAAGGCAAAATGCCGCA
AAAAAGGGAATAAGGGCGACACGGAAATGTTGAATACTCATACTCTTCCTTTTTCAATAT
TATTGAAGCATTTATCAGGGTTATTGTCTCATGAGCGGATACATATTTGAATGTATTTAG
AAAAATAAACAAATAGGGGTTCCGCGCACATTTCCCCGAAAAGTGCCACCTAAATTGTAA
GCGTTAATATTTTGTTAAAATTCGCGTTAAATTTTTGTTAAATCAGCTCATTTTTTAACC
AATAGGCCGAAATCGGCAAAATCCCTTATAAATCAAAAGAATAGACCGAGATAGGGTTGA
GTGTTGTTCCAGTTTGGAACAAGAGTCCACTATTAAAGAACGTGGACTCCAACGTCAAAG
GGCGAAAAACCGTCTATCAGGGCGATGGCCCACTACGTGAACCATCACCCTAATCAAGTT
TTTTGGGGTCGAGGTGCCGTAAAGCACTAAATCGGAACCCTAAAGGGAGCCCCCGATTTA
GAGCTTGACGGGGAAAGCCGGCGAACGTGGCGAGAAAGGAAGGGAAGAAAGCGAAAGGAG
CGGGCGCTAGGGCGCTGGCAAGTGTAGCGGTCACGCTGCGCGTAACCACCACACCCGCCG
CGCTTAATGCGCCGCTACAGGGCGCGTCCCATTCGCCATTCAGGCTGCGCAACTGTTGGG
AAGGGCGATCGGTGCGGGCCTCTTCGCTATTACGCCAGCTGGCGAAAGGGGGATGTGCTG
CAAGGCGATTAAGTTGGGTAACGCCAGGGTTTTCCCAGTCACGACGTTGTAAAACGACGG
CCAGTGAATTGTAATACGACTCACTATAGGGCGAATTGGGTACCGGGCCCCCCCTCGAGG
TCGACGGTATCGATAAGCTTAACTTCATTGTAATCATAATGTTGAAAATTAAACTCTCCC
ATTTTTGTGTTTTTTCTATTTTGTTCACAATTATTTTTATCCATTGAGTCTATATCGTCA
ATGTCAATATTATTTCGATATTTTACATTTTGCAATTCCATTTGTGTAATAGATTGTTCA
CTATGTTTATCTATGTCTTGAGTGTTGCACTCACTTATATTTTCGGTACTATTATATTTT
TCCTTTGTTCTTCCAAGTTTATCATATTCGGTTTCATGAAAAATATTTGCACGGCCATCA
GGAATAATATATTCATAATTATACCCACAATTATTTGTAGTACTTTCACTATTTTGATAA
CTTTCTAATATATTTTTTTTATAATAATTAAGATCATCTTTTTGGGTAATTTCTTTTGAT
GTATCACTTTGTTCTGATATTATACATTTTTTATGTTTGCTATACTGTTTTTTAATATTT
AATGAATTATTTTTATTTTTGCATTTTTTTTTATTATAATACTTATTTATATGATTCTCA
TTAATATTATTTGGGTAATTTTCTGAACAACTTATCGACTCACTATTATATTCATGAATA
TTTTGTGTCAAATTATTGTCGTTTTTCATTTGGTGAATATTATATTTAGAATCTGAATAC
TTACTAGAATTACCGGATTTATAATTATTACTAATATCTATATTTAATACTGGGTGTTTT
TTTGCAAGTTTCCAAGCAATTCTTTTACCTGAGTTTATATTACGCAACTCTTTTATATAT
GGCTTGATTCCTTCTACAAGTTTTGTTAATTCGGGTTCGGGCGCAACAGATAAAGCTCTT
TGTATTACATAATTACCATATTTGTCTAAAATAACATTTTTTAAAACATTTTTTCCTTTT
TTTAGTAGTCCATTTATCATAATTTTTCTACATTTAGTAGTTCCGATTATTAAACATTTT
TCTATAACATTTGATGAAAATTTTTGAACAGCATGCTTTTCAATATCAAGCTTGCATGCC
TGCAG
Restriction sites to linearize plasmid
Partial or complete disruption of the geneComplete
Additional remarks partial/complete disruption
Selectable marker used to select the mutant parasitetgdhfr
Promoter of the selectable markerpbdhfr
Selection (positive) procedurepyrimethamine
Selection (negative) procedureNo
Additional remarks genetic modification
Additional remarks selection procedure
Primer information: Primers used for amplification of the target sequences  Click to view information
Primer information: Primers used for amplification of the target sequences  Click to hide information
Sequence Primer 1AGGATCCGAACAAAACACATTGAAAATTCG
Additional information primer 11514 (BamHI); 5’ targeting region
Sequence Primer 2AGATATCGTTCCTTGCATGTGTGTAG
Additional information primer 21515 (EcoRV); 5’ targeting region
Sequence Primer 3AAAGCTTGATATTGAAAAGCATGCTGTTC
Additional information primer 31522 (HindIII); 3’ targeting region
Sequence Primer 4AAAGCTTAACTTCATTGTAATCATAATGTT
Additional information primer 41523 (HindIII); 3’ targeting region
Sequence Primer 5
Additional information primer 5
Sequence Primer 6
Additional information primer 6