Summary

RMgm-708
Malaria parasiteP. berghei
Genotype
TaggedGene model (rodent): PBANKA_1229000; Gene model (P.falciparum): PF3D7_0801000; Gene product: Plasmodium exported protein (PHISTc), unknown function (PHIST)
Name tag: mCherry
Phenotype Asexual bloodstage;
Last modified: 30 June 2016, 12:58
  *RMgm-708
Successful modificationThe parasite was generated by the genetic modification
The mutant contains the following genetic modification(s) Gene tagging
Reference (PubMed-PMID number) Reference 1 (PMID number) : 23197789
Reference 2 (PMID number) : 27022937
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/ResearcherE.M. Pasini, C.J. Janse, B.M.D. Franke-Fayard
Name Group/DepartmentDepartment of Parasitology
Name InstituteBiomedical Primate Research Centre
CityRijswijk
CountryThe Netherlands
Name of the mutant parasite
RMgm numberRMgm-708
Principal name1921
Alternative name
Standardized name
Is the mutant parasite cloned after genetic modificationNo
Phenotype
Asexual blood stageNo fluorescence was detected. However, IFA analysis using antibodies against mCherry and against the protein showed export into the host erythrocyte with a punctate localisation pattern. (Moreira C. et al., 2016; Plos One 11(3):e0152510).
Gametocyte/GameteNot tested
Fertilization and ookineteNot tested
OocystNot tested
SporozoiteNot tested
Liver stageNot tested
Additional remarks phenotype

Mutant/mutation
The mutant parasite expresses an C-terminal mCherry-tagged version of PBANKA_1229000 (PHIST)

Protein (function)
In the P. berghei genome sequence  3 phist genes (namely, PBANKA_1145400, PBANKA_1229000 and PBANKA_0700800) were identified via BLAST analysis using PHIST domains and pssm profiles as queries. This result differs from the single P. berghei phist gene described in the original annotation. Orthologs of PBANKA_1145400 and PBANKA_122900 were identified in the genome nucleotide sequence databases for other rodent malaria parasites; namely, P. yoelii (PY00289 and PY01786, respectively) and P. chabaudi (PCAS_114490 and PCAS_1229700, respectively). Additional phist genes were not identified in rodent malaria parasites. BLAST analyses using the PHIST domain as queries, identify as reciprocal best hits PBANKA_1229000, the P. vivax PHIST protein PvPHIST/CVC-8195 (PVX_093680), and P. falciparum PF3D7_0801000. This indicates possible orthologous (vertically inherited) relationships and is supported by the observed synteny of adjacent genes, including the ookinete-expressed gene warp, together composing a locus which is conserved across the Plasmodium genus. PBANKA_1229000 is internally localized in the P. berghei genome, but the locus appears to be sub-telomeric in species other than rodent malaria parasites.
The P. berghei PHIST proteins PBANKA_1145400 and PBANKA_1229000 possess similar features to P. falciparum PHIST proteins; namely, signal peptides and PEXEL/HT trafficking motifs; and single PHIST domains, which are divergent in aa sequence with respect to each other. Within the single predicted ORF of PBANKA_0700800 we were unable to identify a signal peptide sequence, and there were no attractive upstream ORFs suggestive of an erroneous gene model. We thus propose that PBANKA_0700800 is a pseudogene. The remaining P. berghei phist genes possess the typical 2-exon gene structure, in which the signal peptide is encoded on the first exon and the PEXEL/HT motif is located within the second exon.

Phenotype
Phenotype analyses of the blood stages of a mutant expressing an mCherry-tagged version of this protein by fluorescence microscopy did not reveal any fluorescence. However, IFA analysis using antibodies against mCherry and against the protein showed export into the host erythrocyte with a punctate localisation pattern (Moreira C. et al., 2016; Plos One 11(3):e0152510).

Additional information
The mutant parasite expresses an mCherry-tagged protein. The protein has been selected for tagging in a screen for putative exported proteins of P. berghei.

The genotype of the parasites has not been analyzed in detail.
Southern analysis of PFG-separated chromosomes (to show integration into the chromosome on which the target gene is located) has been performed (see below). This analysis provided evidence for tagging of the correct gene. The construct used aims at integration of the tagging construct by single-cross-over integration resulting in the presence of a tagged copy of the endogenous gene.




Other mutants
RMgm-718: mutant lacking expression of PBANKA_1229000


  Tagged: Mutant parasite with a tagged gene
Details of the target gene
Gene Model of Rodent Parasite PBANKA_1229000
Gene Model P. falciparum ortholog PF3D7_0801000
Gene productPlasmodium exported protein (PHISTc), unknown function
Gene product: Alternative namePHIST
Details of the genetic modification
Name of the tagmCherry
Details of taggingC-terminal
Additional remarks: tagging
Commercial source of tag-antibodiesClontech
Type of plasmid/construct(Linear) plasmid single 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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CTAAATTGTAAGCGTTAATATTTTGTTAAAATTCGCGTTAAATTTTTGTTAAATCAGCTC
ATTTTTTAACCAATAGGCCGAAATCGGCAAAATCCCTTATAAATCAAAAGAATAGACCGA
GATAGGGTTGAGTGTTGTTCCAGTTTGGAACAAGAGTCCACTATTAAAGAACGTGGACTC
CAACGTCAAAGGGCGAAAAACCGTCTATCAGGGCGATGGCCCACTACGTGAACCATCACC
CTAATCAAGTTTTTTGGGGTCGAGGTGCCGTAAAGCACTAAATCGGAACCCTAAAGGGAG
CCCCCGATTTAGAGCTTGACGGGGAAAGCCGGCGAACGTGGCGAGAAAGGAAGGGAAGAA
AGCGAAAGGAGCGGGCGCTAGGGCGCTGGCAAGTGTAGCGGTCACGCTGCGCGTAACCAC
CACACCCGCCGCGCTTAATGCGCCGCTACAGGGCGCGTCCCATTCGCCATTCAGGCTGCG
CAACTGTTGGGAAGGGCGATCGGTGCGGGCCTCTTCGCTATTACGCCAGCTGGCGAAAGG
GGGATGTGCTGCAAGGCGATTAAGTTGGGTAACGCCAGGGTTTTCCCAGTCACGACGTTG
TAAAACGACGGCCAGTGAGCGCGCGTAATACGACTCACTATAGGGCGAATTGGAGCTCCA
CCGCGGTGGCGGCCGCTCTAGAACTAGTGAGTATGGGAATATGGGATGGTTTGAAACCTT
CTTCAATGAGCAGAAGTCAAGGAATGACCATAAATCAATATCCATCAACAAGTGGAACCC
AAGGAATAACCATAAATCAATATCCATCAACAAGTGGAACCCAAGGAATGACCAGAAATC
AATATCCATCAACAAGTGGAACCCAAGGAATGACCAGAAATCAATATCCATCAACAAGTG
GAACCCAAGGAATGACCAGAAATCAATATCCATCAACAAGTGGAACCCAAGGAATGACCA
GAAATCAATATCCATCAACAAGTGGAACCCAAGGAATGACCAGAAATCAATATCCATCAA
CAAGTGGAACCCAAGGAATGACCAGAAATCAATTAATTGGAGAGATGTATAATATTGATG
AGCGTAAAATAGAAGAACCAATGTGGTTAGGTTTGAAAAGCTCATATAAACCTACTCCAA
GAAAACCATATGCATCAAACTTTTCTGATATGGAAACATTCCCACGAGAAAATTATTCAA
GAAATAAACATGAGGCATTTGATAGATTCGATTCACACAATATGCCAAGAGGTCATAGTA
CTAAATACCCTACACGCACATCTTCAGAATTTCATAATCCAATGAATAGATATAATTCTA
GGGGCACTTCAAACTCAGGAAGTATATTAAATTCAATGAAAAGTTTACGAGGAAATGATT
CTCATAGACTAGATAATGGACAAAATTATATGAGTGAACCAATATCTACAACATCCATGG
GAAACCATAAATTATATTTTGGAAGTAAAAATACTCAAGAAACTATAAGTACATCATTCA
AATCTAAATCAAATGATTATAAGGGTCTTGATCAACAGAATAAAGCTAGTAATTATAATT
CAGATACAGATGCAGTTATAGATAGTAAAGAAGTAAGTGAAACTAATTTTAGTCCAAGAA
AGTCACCAGATGCTCAAATCAAAGAAATGAATGATCATATAAATGAAAAAATTGACATGT
TAGATGAAAATGCAACTCCAGATGTATTTCGTAATATTTGGACAATTTTCATTTCTGCTG
AAACTAAAAAATTGATGATGACTCAAGAATATGTATTACAATATTCTATATATCTACAAA
AACGAAATAAATTATATCCAGAAATGCGCACGGCGGCATGGTGGAAAGTACATTATTCTA
TGATTAATAAATTTATAAAGCGTGAAAAAGAAGAGGTTTCTGAATTAAAAGAATTATTAA
AATCATCTACTAATACATATTCTAATTTCGTCACGTTTATTAGAAATAAAATGGAGTCTT
GGAAAGAATTCCAGAAAGAAATTAAAGATACATGGTTGGCAACTTTAACATACAAAATTA
ATAAATACAGTTCTCAAGACAATTATGGATCCGGTGGAGGTGGAAGATCCATGGTGAGCA
AGGGCGAGGAGGATAACATGGCCATCATCAAGGAGTTCATGCGCTTCAAGGTGCACATGG
AGGGCTCCGTGAACGGCCACGAGTTCGAGATCGAGGGCGAGGGCGAGGGCCGCCCCTACG
AGGGCACCCAGACCGCCAAGCTGAAGGTGACCAAGGGTGGCCCCCTGCCCTTCGCCTGGG
ACATCCTGTCCCCTCAGTTCATGTACGGCTCCAAGGCCTACGTGAAGCACCCCGCCGACA
TCCCCGACTACTTGAAGCTGTCCTTCCCCGAGGGCTTCAAGTGGGAGCGCGTGATGAACT
TCGAGGACGGCGGCGTGGTGACCGTGACCCAGGACTCCTCCCTGCAGGACGGCGAGTTCA
TCTACAAGGTGAAGCTGCGCGGCACCAACTTCCCCTCCGACGGCCCCGTAATGCAGAAGA
AGACCATGGGCTGGGAGGCCTCCTCCGAGCGGATGTACCCCGAGGACGGCGCCCTGAAGG
GCGAGATCAAGCAGAGGCTGAAGCTGAAGGACGGCGGCCACTACGACGCTGAGGTCAAGA
CCACCTACAAGGCCAAGAAGCCCGTGCAGCTGCCCGGCGCCTACAACGTCAACATCAAGT
TGGACATCACCTCCCACAACGAGGACTACACCATCGTGGAACAGTACGAACGCGCCGAGG
GCCGCCACTCCACCGGCGGCATGGACGAGCTGTACAAGTAATCTAGAATTATTAATATAT
ATGAATATATATACATCGTTGTATGCGTAATATACGTGTAACTGTTTTTTTTAATTTTTA
TATTAACTTTTCCTTATACATTTTAATGCCATTAACTAATAATATTTTACTTGAAAAAAT
TCTAAATTTTTATAACTCTAATATAAAATCTGTGAATAATAAAAAATTAGCATAAAGTAC
GAATTAAACATCGAATATTTGTTTTAAAGGGGCACACATTATGTGCATATAATTTTATTC
TTTGAATTTAATTTTTTGCTTATAATTAGATTAAATTAATATAATTTTTTTATTGAAAAT
AAAAACTTATTTTAAAAAATTAACAAATGGAAGTTAATGAAAAATACGAAAAAAGTAAAG
GGTTAATTCTTATATGGTCGGTACCGAGCTCGAATTCGATATCGAAATTGAAGGAAAAAA
CATCATTTGTGTTTATATGTTTTTTTTTAATTTTTCAACTTTTTTTATTTAACTATGCTA
ATCATATTATAAACATTTTAAAAATCAAGTCCAACTATTTATGAATCATTGAAGAGACAA
CATAATTATAACAGTAAAAATAATAATAATAATAATAATTTTGTCTTATTGTTTCAAACA
TATTATGTTCCATAAATATCTAAATTTTTAATATGGTCATAAATTCATAGGATTTAATTT
ATATAAAATATGGCTATTCATACTAGCCATTTTATGTGTGTAAAGTTTTTTTTTTCTGAA
CAAAAAATAAAATGTTCATAATTTTAGCTATTTACATGCATGTGCATGCACAAAAAAAAA
TATGCACACAACATACACATTTTTACAGTTATAAATACAATCAATTGGTATAAATATATA
AGTAAGAAAAACGGGATCAAAGCTAGCTTCTGTATTTCCGCTAGACAGCCATCTCCATCT
GGATTCGTCCGTGCGGGACGTAGCCCACGACCTCAAAATCCTCGGCGGTGAAATCGTCGA
TTTCCTTGATGCGTTCCTTGTTGAGGATGTTCACAATGGGGAACGGTCTCGGTTCTCTCC
GCAGCTGCTCTTTTAAAGCCTCGACATGGTTCGTGTAGACATGCGTGTTCCCCATGAAGT
GAATGAACTCCTTAGGTTTTAGGTTGCAGACGTGTGCAACCATGAGCGTCAAAAGCGAAT
AGGAAGCGATGTTGAAGGGGACGCCGAGGCCGACATCGCACGACCGCTGATACATGATGC
ACGACAGCTCCTTCTGGTCGTTCACGTAGAACTGGCACAACAAGTGACAAGGCGGCAGCG
CCATTTCGTCCAGCGCTGCAGGATTCCAGGCAGTCATGAGCATGCGACGATCTGTTGGAT
TCGTTCTCAGCATCTGGATCACATTCTTCAGCTGGTCGACGCCCTGCCCTGTGTAGTCTG
TGTGCATGTCTTTGTATGCCGCGCCGAAGTGTCTCCACTGGAAGCCGTAGCCCGGGCCGA
TGTCTCCGACCTCTCGGTGGGGGAGATTGCGCGAATCGAGGAACTCGCGTGTCACATTCT
TGTCCCAGATCTTCACGCCCTTCTCAGAAAGATGGTTTGCGTTCGTGTCGCCGCGAATGA
ACCACAGCAACTCTTCGAGGACCCCTTTCCAGAACACACGCTTTGTGGTGAGAAGTGGAA
AGGCCTGATCCAGCGAGTAGCGCATAGTGCAGCCGAATTTGGAGATGACACCAACGCCCG
TTCGGTCATCCATTGTCCTTCCATTGTTAATAATGTCGGCAATGAGATCAAGGTACTGGA
ACTCTTCATGGCCTCTAAAGTGAACATGCGGAACGGCCCGAATCAGTTCCTTTTGCTCGC
GTTTTTTCCGGTCTTCTTCGTCCATCCACGCCAACACCGGGGCAATGGCTGCGGCCGAAG
AAGGAGCCTGCAACCCGTGCACGGGAGTTGTCTCCCTCGTGGACGTCAAGGAGCTCATTG
CGTTGCTCGGTTCCGCAGTGGCTGCGTCGTCAGTCTTCCTTCTCTTCTCGAGAACCACAA
AGTCGTAGGGAACCCCGTTGTCTGAGAAGGTCTTGGAAATGAAGATGGGTCGATACGTCG
CTTCATTGTCCTTCTCTCTTCCGAGCTCCGGACAAAAGGGAACGAACACAGACTCGGCAG
GAGCTGCAGCCTGCGCAGCAGTTGATTTGTTTGAAAGAATGTCATCTCCGGGGAACGCAG
GGAAGAAAACGTCGCACGGAAACTCGCGGGCTACACGCGTGATGTACAGGTGAGAGGCAA
CGCCCAGAGACAGCGCTGCCTCGTACAGTCCCGCTCCTCCCACGACAAAAATCTGGTCGA
CAGAATCCTTGTACTCTTCCTCCAGAAGGCTGAGAGCTGCTGGGAGTGAAGCACAGACTC
GGACGCGCTGCTGGCCTTCAGCTTGAGGCTTCTCCGCCGCAATGTCTTCTTCTTTGAGGG
AAGAGGAAACGACGATGTTCAATCTGTCCACGAGGGGTCTAAACTTTCGAGGCATGCTTT
CCCAGGTTTTCCGTCCCATGACAACGGCGTTGAATCTCTTGCCGACTGATGGAGAGGGAA
GTCCAGAGTCGCCCGTCTTTGCAAATTTCCTGGGAAGCCACCCGTTCAGGCGACTGGCTT
CTTCGGGCGTCGTTTTTGTCACACGAGAAAAGTGTTTGAAATCTGTGGTCAAGTGGGGCC
ACGGGAGGCCGTTGTTGATGCCGATGCCCCTCTTGGGGGTCATCGCGACGACCAGACACA
CCGGTTTATGCATTGATCCTTTGTAACATTTAGGTGTGTATTTATATATATAAGCAATAT
ACAAATGAAAATACTATTATAAAACGGAAACTCAAAAAATGTTGAAACAAATAGTCAATT
TTTAAAATGAATGTGTAAATTGTTTAAGTTTTATGACAAAAAAAGGAAAAGTGTATATAA
AATAAAAATTGGAAAAAATTTATATATTACATGCCATATAATACCATTTATTTACTATTT
ATATCTTAATATAAAAAATTGTAGCAAAAATATATAATCGTATTTTATATATTTAATATT
ATATAACCCCTTAAAAAAAAAAATTTAATTGAAAAAATTTTAATTTATGCACTTTTTTAT
TGTATTATATATATTCCGTTATTTTAATTACACATAAACTTAATAGAAAAATTAATTCGC
TGCTAAATAAAAATTATAGGGCTATATATGATGGGATCATATTCTATCTTTCTCTTTTTC
AAATYCTTTATATTTTAGAATACAAATCGCTTTAAACAATAAAAAAGAGGTATATCCTAC
CCGAATTGACACACACAATAAATAATAAATATATAAATAATAAAAATATAAATATATAAA
TTATAAATATATAAATTATATATATAAATATATTAACATGTGTGTAATTTGTTAAAATTG
ATATTAAACAGCTTTGCATGGGGAAAGAGTGGCTTTTATAAGCGAATCATATAATACAGT
GTTAATTTTAATATATAAAATATAATTTCCTTAAATTGTTATTTAACATTTTTAATTGCC
TACCCARGGTAATATAATATCCAAATGACAATCAAAGATAAATGAAATACCGCTCCATTT
TTCCTATATATCCATTTTCATTTTGTGTAAATGTTCATAAAATTTTATCATATACTCATA
AAAAACAGGGGAAAAATGAAAAAATGAACGTCCACCCCGTGTGAATATGCTCATTTTGTA
TACTATACATTTTTTTTTATATAATTTATATTTTCCAAATAAAAAAAATGTTTGTACATG
TATATATCAATATTGTGTTATGGCATGATGAAAAACAATTATTTTATTCGTTTTTAAATT
TGTAGAATTTCCCTAAATTATTAATGATTTCATATTATAATATATGTATCTATATATTTT
TAATGTGTAATTCAAAATGGCGAAAAAGAAACAAACCATACTTTATAAGCAACTCTCTTT
GAGCACTTACCATAAAAAATTATACTACTAAATTCAAATTAAATAAAAAAATATATATTA
TAAAATTGGCTTTAATTATTTATAAGCACTTGCTTTGTTTTACTCAAAAAGTATTGGCAT
AAGTATAAACATAAAACCAATTTTTCACAAAATATATTATATGTCCATACAACTATATCC
GAACYTTCAAAACACATCCAAATTTCCATTTACCACATGCAAGCAAAAGGATATTAAATA
AAAAAAAATTATTTATATTTTCATGATCACTTAAATAATATTTAAAAAAAAAAGAGACCA
TATATAAAATGTATAATTTAAATATAGCTAACTTATTAAAAAAAATGAATATTGAGCATA
TATCCATAATTTAAAACACTTCTATTTTATGTTATTTGATATATAAGTCGATAAATTATT
GCATGTATGCATATAGACAGATTAATATACACCAAAATAATAAAAATTATATATAGTTCA
TATATTATTCTGAATAATAAATTAGCTTTCCTCTTAATTCACTTGGTATGTATATATATA
TATATATATATATATATATATATATGGATAAATAAATATAAGGTCTATAACAGGAAAAAA
TGTGTATTAACTTTTTTTTATAATTATTTTATATTTTTTCTCCATTTATTTAAATATGTT
AAAATTATTTCCCATTTTATAAACAAAAATTAATCCATAAAATAGCAAAATAACAAAATG
ACAAAATAACAAAAACAAAAAAACGCATTATATGAGTTCATTTTACACAATCCAAAGAAA
AAGAAAATATGCTTAAAATATTCATAACACACTTTTAAGCTATAAAAATATATATATTTA
TTTGTGTCTATATTACCAACTCAATTTAATAGATGTGTTATGTGATTAATTCATACACAA
ACATACAAAAATAAACACCATTTTTGAAAAGATTAATTTGAAATCATATATGTTATTATT
ATATATTTTTATATCAATGAAATTTAAAAAAATAAAAGGGAAATCAATGTATTAAAAATA
ATTATATGTTATTTTATTTCCACAATATTTATTTATTATTTATTGTTGACCTGCAGGCAT
GCAAGCTAGCTTATCGATACCGTCGACCTCGAGGGGGGGCCCGGTACCCAGCTTTTGTTC
CCTTTAGTGAGGGTTAATTGCGCGCTTGGCGTAATCATGGTCATAGCTGTTTCCTGTGTG
AAATTGTTATCCGCTCACAATTCCACACAACATACGAGCCGGAAGCATAAAGTGTAAAGC
CTGGGGTGCCTAATGAGTGAGCTAACTCACATTAATTGCGTTGCGCTCACTGCCCGCTTT
CCAGTCGGGAAACCTGTCGTGCCAGCTGCATTAATGAATCGGCCAACGCGCGGGGAGAGG
CGGTTTGCGTATTGGGCGCTCTTCCGCTTCCTCGCTCACTGACTCGCTGCGCTCGGTCGT
TCGGCTGCGGCGAGCGGTATCAGCTCACTCAAAGGCGGTAATACGGTTATCCACAGAATC
AGGGGATAACGCAGGAAAGAACATGTGAGCAAAAGGCCAGCAAAAGGCCAGGAACCGTAA
AAAGGCCGCGTTGCTGGCGTTTTTCCATAGGCTCCGCCCCCCTGACGAGCATCACAAAAA
TCGACGCTCAAGTCAGAGGTGGCGAAACCCGACAGGACTATAAAGATACCAGGCGTTTCC
CCCTGGAAGCTCCCTCGTGCGCTCTCCTGTTCCGACCCTGCCGCTTACCGGATACCTGTC
CGCCTTTCTCCCTTCGGGAAGCGTGGCGCTTTCTCATAGCTCACGCTGTAGGTATCTCAG
TTCGGTGTAGGTCGTTCGCTCCAAGCTGGGCTGTGTGCACGAACCCCCCGTTCAGCCCGA
CCGCTGCGCCTTATCCGGTAACTATCGTCTTGAGTCCAACCCGGTAAGACACGACTTATC
GCCACTGGCAGCAGCCACTGGTAACAGGATTAGCAGAGCGAGGTATGTAGGCGGTGCTAC
AGAGTTCTTGAAGTGGTGGCCTAACTACGGCTACACTAGAAGGACAGTATTTGGTATCTG
CGCTCTGCTGAAGCCAGTTACCTTCGGAAAAAGAGTTGGTAGCTCTTGATCCGGCAAACA
AACCACCGCTGGTAGCGGTGGTTTTTTTGTTTGCAAGCAGCAGATTACGCGCAGAAAAAA
AGGATCTCAAGAAGATCCTTTGATCTTTTCTACGGGGTCTGACGCTCAGTGGAACGAAAA
CTCACGTTAAGGGATTTTGGTCATGAGATTATCAAAAAGGATCTTCACCTAGATCCTTTT
AAATTAAAAATGAAGTTTTAAATCAATCTAAAGTATATATGAGTAAACTTGGTCTGACAG
TTACCAATGCTTAATCAGTGAGGCACCTATCTCAGCGATCTGTCTATTTCGTTCATCCAT
AGTTGCCTGACTCCCCGTCGTGTAGATAACTACGATACGGGAGGGCTTACCATCTGGCCC
CAGTGCTGCAATGATACCGCGAGACCCACGCTCACCGGCTCCAGATTTATCAGCAATAAA
CCAGCCAGCCGGAAGGGCCGAGCGCAGAAGTGGTCCTGCAACTTTATCCGCCTCCATCCA
GTCTATTAATTGTTGCCGGGAAGCTAGAGTAAGTAGTTCGCCAGTTAATAGTTTGCGCAA
CGTTGTTGCCATTGCTACAGGCATCGTGGTGTCACGCTCGTCGTTTGGTATGGCTTCATT
CAGCTCCGGTTCCCAACGATCAAGGCGAGTTACATGATCCCCCATGTTGTGCAAAAAAGC
GGTTAGCTCCTTCGGTCCTCCGATCGTTGTCAGAAGTAAGTTGGCCGCAGTGTTATCACT
CATGGTTATGGCAGCACTGCATAATTCTCTTACTGTCATGCCATCCGTAAGATGCTTTTC
TGTGACTGGTGAGTACTCAACCAAGTCATTCTGAGAATAGTGTATGCGGCGACCGAGTTG
CTCTTGCCCGGCGTCAATACGGGATAATACCGCGCCACATAGCAGAACTTTAAAAGTGCT
CATCATTGGAAAACGTTCTTCGGGGCGAAAACTCTCAAGGATCTTACCGCTGTTGAGATC
CAGTTCGATGTAACCCACTCGTGCACCCAACTGATCTTCAGCATCTTTTACTTTCACCAG
CGTTTCTGGGTGAGCAAAAACAGGAAGGCAAAATGCCGCAAAAAAGGGAATAAGGGCGAC
ACGGAAATGTTGAATACTCATACTCTTCCTTTTTCAATATTATTGAAGCATTTATCAGGG
TTATTGTCTCATGAGCGGATACATATTTGAATGTATTTAGAAAAATAAACAAATAGGGGT
TCCGCGCACATTTCCCCGAAAAGTGCCAC
Restriction sites to linearize plasmid NdeI
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 1AACTAGTGAGTATGGGAATATGGGATG
Additional information primer 1L6226 (SpeI); PBANKA_122900 Forward
Sequence Primer 2AGGATCCATAATTGTCTTGAGAACTGTATTTATTAATTTTG
Additional information primer 2L6227 (BamHI); PBANKA_122900 Reverse
Sequence Primer 3
Additional information primer 3
Sequence Primer 4
Additional information primer 4
Sequence Primer 5
Additional information primer 5
Sequence Primer 6
Additional information primer 6