Summary

RMgm-31
Malaria parasiteP. berghei
Genotype
TaggedGene model (rodent): PBANKA_1444000; Gene model (P.falciparum): PF3D7_1229400; Gene product: macrophage migration inhibitory factor (MIF)
Name tag: c-myc
Phenotype Asexual bloodstage; Gametocyte/Gamete;
Last modified: 20 August 2012, 12:51
  *RMgm-31
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) : 17158894
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/ResearcherK.D. Augustijn, C.J. Janse, A.P. Waters
Name Group/DepartmentLeiden Malaria Research Group
Name InstituteLeiden University Medical Center
CityLeiden
CountryThe Netherlands
Name of the mutant parasite
RMgm numberRMgm-31
Principal name618cl1
Alternative namepbMIF-MYC
Standardized name
Is the mutant parasite cloned after genetic modificationYes
Phenotype
Asexual blood stageExpression of the c-myc-tagged protein (MIF-MYC) throughout asexual stages; IFA analysis using c-myc antibodies showed low staining in early ring forms. Staining in trophozoites and schizonts appeared to be restricted to parasitophorous vacuole.
Gametocyte/GameteNormal gametocytes; gametocytes expressed the c-myc tagged proteins as shown by IFA analysis using c-myc antibodies
Fertilization and ookineteNot tested
OocystNot tested
SporozoiteNot tested
Liver stageNot tested
Additional remarks phenotype

Mutant/mutation
The mutant expresses a c-myc-tagged (c-terminal) form of Macrophage Migration Inhibitory Factor protein (MIF-MYC).

Protein (function)
Macrophage migration inhibitory factor (MIF) is a mammalian cytokine that participates in innate and adaptive immune responses. Homologues of mammalian MIF have been discovered in parasite species (nematodes and malaria parasites). Like human MIF, histidine-tagged purified recombinant Plasmodium MIF shows tautomerase and oxidoreductase activities (although the activities are reduced compared to those of histidine-tagged human MIF) and efficiently inhibits AP-1 activity in human embryonic kidney cells (Augustijn et al., Infect Immun. (2007),75:1116-28). P. berghei MIF is expressed in both the mammalian host and mosquito vector. In blood stages, MIF is secreted into the infected erythrocytes and released upon schizont rupture.

Phenotype
MIF-MYC is expressed in all blood stages. MIF-MYC expression has not been analysed during mosquito and liver stage development.

Other mutants 
RMgm-27 is a transgenic mutant expressing GFP-tagged MIF.
RMgm-26 is a mutant lacking expression of MIF
RMgm-665: A P. yoelii (17XNL) mutant lacking expression of MIF
RMgm-582: A P.yoelii (Py17X) mutant containing  2 additional copies of the macrophage migration inhibitory factor gene (mif) of P. yoelii integrated into the c-ssu-rrna gene locus. These mif copies are under control of the strong consecutive eef1a promoter.
RMgm-583: A P. yoelii (17XL) mutant containing one additional copy of the macrophage migration inhibitory factor gene (mif) of P. yoelii integrated into the dhfr/ts gene locus


  Tagged: Mutant parasite with a tagged gene
Details of the target gene
Gene Model of Rodent Parasite PBANKA_1444000
Gene Model P. falciparum ortholog PF3D7_1229400
Gene productmacrophage migration inhibitory factor
Gene product: Alternative nameMIF
Details of the genetic modification
Name of the tagc-myc
Details of taggingC-terminal
Additional remarks: tagging
Commercial source of tag-antibodiesSigma C3956 0.2 ml
Type of plasmid/constructPlasmid 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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GGGCCCCCCCTCGAGGTCGACGGTATCGATAAGCTTGCATGCCTGCAGGTCAACAATAAA
TAATAAATAAATATTGTGGAAATAAAATAACATATAATTATTTTTAATACATTGATTTCC
CTTTTATTTTTTTAAATTTCATTGATATAAAAATATATAATAATAACATATATGATTTCA
AATTAATCTTTTCAAAAATGGTGTTTATTTTTGTATGTTTGTGTATGAATTAATCACATA
ACACATCTATTAAATTGAGTTGGTAATATAGACACAAATAAATATATATATTTTTATAGC
TTAAAAGTGTGTTATGAATATTTTAAGCATATTTTCTTTTTCTTTGGATTGTGTAAAATG
AACTCATATAATGCGTTTTTTTGTTTTTGTTATTTTGTCATTTTGTTATTTTGCTATTTT
ATGGATTAATTTTTGTTTATAAAATGGGAAATAATTTTAACATATTTAAATAAATGGAGA
AAAAATATAAAATAATTATAAAAAAAAGTTAATACACATTTTTTCCTGTTATAGACCTTA
TATTTATTTATCCATATATATATATATATATATATATATATATATATACATACCAAGTGA
ATTAAGAGGAAAGCTAATTTATTATTCAGAATAATATATGAACTATATATAATTTTTATT
ATTTTGGTGTATATTAATCTGTCTATATGCATACATGCAATAATTTATCGACTTATATAT
CAAATAACATAAAATAGAAGTGTTTTAAATTATGGATATATGCTCAATATTCATTTTTTT
TAATAAGTTAGCTATATTTAAATTATACATTTTATATATGGTCTCTTTTTTTTTTAAATA
TTATTTAAGTGATCATGAAAATATAAATAATTTTTTTTTATTTAATATCCTTTTGCTTGC
ATGTGGTAAATGGAAATTTGGATGTGTTTTGAARGTTCGGATATAGTTGTATGGACATAT
AATATATTTTGTGAAAAATTGGTTTTATGTTTATACTTATGCCAATACTTTTTGAGTAAA
ACAAAGCAAGTGCTTATAAATAATTAAAGCCAATTTTATAATATATATTTTTTTATTTAA
TTTGAATTTAGTAGTATAATTTTTTATGGTAAGTGCTCAAAGAGAGTTGCTTATAAAGTA
TGGTTTGTTTCTTTTTCGCCATTTTGAATTACACATTAAAAATATATAGATACATATATT
ATAATATGAAATCATTAATAATTTAGGGAAATTCTACAAATTTAAAAACGAATAAAATAA
TTGTTTTTCATCATGCCATAACACAATATTGATATATACATGTACAAACATTTTTTTTAT
TTGGAAAATATAAATTATATAAAAAAAAATGTATAGTATACAAAATGAGCATATTCACAC
GGGGTGGACGTTCATTTTTTCATTTTTCCCCTGTTTTTTATGAGTATATGATAAAATTTT
ATGAACATTTACACAAAATGAAAATGGATATATAGGAAAAATGGAGCGGTATTTCATTTA
TCTTTGATTGTCATTTGGATATTATATTACCYTGGGTAGGCAATTAAAAATGTTAAATAA
CAATTTAAGGAAATTATATTTTATATATTAAAATTAACACTGTATTATATGATTCGCTTA
TAAAAGCCACTCTTTCCCCATGCAAAGCTGTTTAATATCAATTTTAACAAATTACACACA
TGTTAATATATTTATATATATAATTTATATATTTATAATTTATATATTTATATTTTTATT
ATTTATATATTTATTATTTATTGTGTGTGTCAATTCGGGTAGGATATACCTCTTTTTTAT
TGTTTAAAGCGATTTGTATTCTAAAATATAAAGRATTTGAAAAAGAGAAAGATAGAATAT
GATCCCATCATATATAGCCCTATAATTTTTATTTAGCAGCGAATTAATTTTTCTATTAAG
TTTATGTGTAATTAAAATAACGGAATATATATAATACAATAAAAAAGTGCATAAATTAAA
ATTTTTTCAATTAAATTTTTTTTTTTAAGGGGTTATATAATATTAAATATATAAAATACG
ATTATATATTTTTGCTACAATTTTTTATATTAAGATATAAATAGTAAATAAATGGTATTA
TATGGCATGTAATATATAAATTTTTTCCAATTTTTATTTTATATACACTTTTCCTTTTTT
TGTCATAAAACTTAAACAATTTACACATTCATTTTAAAAATTGACTATTTGTTTCAACAT
TTTTTGAGTTTCCGTTTTATAATAGTATTTTCATTTGTATATTGCTTATATATATAAATA
CACACCTAAATGTTACAAAGGATCAATGCATAAACCGGTGTGTCTGGTCGTCGCGATGAC
CCCCAAGAGGGGCATCGGCATCAACAACGGCCTCCCGTGGCCCCACTTGACCACAGATTT
CAAACACTTTTCTCGTGTGACAAAAACGACGCCCGAAGAAGCCAGTCGCCTGAACGGGTG
GCTTCCCAGGAAATTTGCAAAGACGGGCGACTCTGGACTTCCCTCTCCATCAGTCGGCAA
GAGATTCAACGCCGTTGTCATGGGACGGAAAACCTGGGAAAGCATGCCTCGAAAGTTTAG
ACCCCTCGTGGACAGATTGAACATCGTCGTTTCCTCTTCCCTCAAAGAAGAAGACATTGC
GGCGGAGAAGCCTCAAGCTGAAGGCCAGCAGCGCGTCCGAGTCTGTGCTTCACTCCCAGC
AGCTCTCAGCCTTCTGGAGGAAGAGTACAAGGATTCTGTCGACCAGATTTTTGTCGTGGG
AGGAGCGGGACTGTACGAGGCAGCGCTGTCTCTGGGCGTTGCCTCTCACCTGTACATCAC
GCGTGTAGCCCGCGAGTTTCCGTGCGACGTTTTCTTCCCTGCGTTCCCCGGAGATGACAT
TCTTTCAAACAAATCAACTGCTGCGCAGGCTGCAGCTCCTGCCGAGTCTGTGTTCGTTCC
CTTTTGTCCGGAGCTCGGAAGAGAGAAGGACAATGAAGCGACGTATCGACCCATCTTCAT
TTCCAAGACCTTCTCAGACAACGGGGTTCCCTACGACTTTGTGGTTCTCGAGAAGAGAAG
GAAGACTGACGACGCAGCCACTGCGGAACCGAGCAACGCAATGAGCTCCTTGACGTCCAC
GAGGGAGACAACTCCCGTGCACGGGTTGCAGGCTCCTTCTTCGGCCGCAGCCATTGCCCC
GGTGTTGGCGTGGATGGACGAAGAAGACCGGAAAAAACGCGAGCAAAAGGAACTGATTCG
GGCCGTTCCGCATGTTCACTTTAGAGGCCATGAAGAGTTCCAGTACCTTGATCTCATTGC
CGACATTATTAACAATGGAAGGACAATGGATGACCGAACGGGCGTTGGTGTCATCTCCAA
ATTCGGCTGCACTATGCGCTACTCGCTGGATCAGGCCTTTCCACTTCTCACCACAAAGCG
TGTGTTCTGGAAAGGGGTCCTCGAAGAGTTGCTGTGGTTCATTCGCGGCGACACGAACGC
AAACCATCTTTCTGAGAAGGGCGTGAAGATCTGGGACAAGAATGTGACACGCGAGTTCCT
CGATTCGCGCAATCTCCCCCACCGAGAGGTCGGAGACATCGGCCCGGGCTACGGCTTCCA
GTGGAGACACTTCGGCGCGGCATACAAAGACATGCACACAGACTACACAGGGCAGGGCGT
CGACCAGCTGAAGAATGTGATCCAGATGCTGAGAACGAATCCAACAGATCGTCGCATGCT
CATGACTGCCTGGAATCCTGCAGCGCTGGACGAAATGGCGCTGCCGCCTTGTCACTTGTT
GTGCCAGTTCTACGTGAACGACCAGAAGGAGCTGTCGTGCATCATGTATCAGCGGTCGTG
CGATGTCGGCCTCGGCGTCCCCTTCAACATCGCTTCCTATTCGCTTTTGACGCTCATGGT
TGCACACGTCTGCAACCTAAAACCTAAGGAGTTCATTCACTTCATGGGGAACACGCATGT
CTACACGAACCATGTCGAGGCTTTAAAAGAGCAGCTGCGGAGAGAACCGAGACCGTTCCC
CATTGTGAACATCCTCAACAAGGAACGCATCAAGGAAATCGACGATTTCACCGCCGAGGA
TTTTGAGGTCGTGGGCTACGTCCCGCACGGACGAATCCAGATGGAGATGGCTGTCTAGCG
GAAATACAGAAGCTAGCTTTGATCCCGTTTTTCTTACTTATATATTTATACCAATTGATT
GTATTTATAACTGTAAAAATGTGTATGTTGTGTGCATATTTTTTTTTGTGCATGCACATG
CATGTAAATAGCTAAAATTATGAACATTTTATTTTTTGTTCAGAAAAAAAAAACTTTACA
CACATAAAATGGCTAGTATGAATAGCCATATTTTATATAAATTAAATCCTATGAATTTAT
GACCATATTAAAAATTTAGATATTTATGGAACATAATATGTTTGAAACAATAAGACAAAA
TTATTATTATTATTATTATTTTTACTGTTATAATTATGTTGTCTCTTCAATGATTCATAA
ATAGTTGGACTTGATTTTTAAAATGTTTATAATATGATTAGCATAGTTAAATAAAAAAAG
TTGAAAAATTAAAAAAAAACATATAAACACAAATGATGTTTTTTCCTTCAATTTCGATAT
CCGCATATCGAAATGATGCTATCATTTTTTATGGAGTATTGAATATGTGATAAACACAAG
TTGAGAGGGGCGTGCCGAAATTAAATAAAATCCTCTCAAAATTGATGAGAAGTGTATTAT
ACTATGCACATGCTTATTTACGAGTACATGCCTAAGCGCTATTTGCTAAATTTTGTGATT
TTTCATAAGAAAATTTATAGTTTGTGTGTGGAGAATATTACTAAAATACATAGAAAGCTG
AATGAGAAAACATGGAAGAAAAATAAAAAAAAGAAGATAAATAGCAATAAAAAATATTCT
CTAAACACATCGATTTTAACTACAATATTAAAAGGGTATTTAATTAATTTATTTTTTGTT
TTAAGTATTCGAATTCAGTATTGCACGACTAGATATTTGTTTTATATTTTTTCAAATAGC
AAATTATTTTATTTCTCCTTAAATTGATATATGGAAAAAAATATATAAATAAAAAATAAC
AGCAATAATAATAAACATTTGCCTTACATAATAGTAATATGCAATATATAGCTATATGTA
ATGGTGAGTGAAAATCAGTTAAATGTGGGGTAAAAAAGAGTGTATATACCTTTTTTTTAA
AAAGAAAAGTTTTATCTGTGCAAAAAAAAAAATTAATTAAAATTATTAATATATGTGAAT
GTTAAGCATATGCGGTGTGAAATACCGCACAGATGCGTAAGGAGAAAATACCGCATCAGG
CGCCATTCGCCATTCAGGCTGCGCAACTGTTGGGAAGGGCGATCGGTGCGGGCCTCTTCG
CTATTACGCCAGCTGGCGAAAGGGGGATGTGCTGCAAGGCGATTAAGTTGGGTAACGCCA
GGGTTTTCCCAGTCACGACGTTGTAAAACGACGGCCAGTGAATTCGAAATTGAAGGAAAA
AACATCATTTGTGTTTATATGTTTTTTTTTAATTTTTCAACTTTTTTTATTTAACTATGC
TAATCACATTATAAACATTTTAAAAATCAAGTCCAACTATTTATGAATCATTGAAGAGAC
AACATAATTATAACAGTAAAAATAATAATAATAATAATAATTTTGTCTTATTGTTTCAAA
CATATTATGTTCCATAAATATCTAAATTTTTAATATGGTCATAAATTCATAGGATTTAAT
TTATATAAAATATGGCTATTCATACTAGCCATTTTATGTGTGTAAAGTTTTTTTTTTCTG
AACAAAAAATAAAATGTTCATAATTTTAGCTATTTACATGCATGTGCATGCACAAAAAAA
AATATGCACACAACATACACATTTTTACAGTTATAAATACAATCAATTGGTATAAATATA
TAAGTAAGAAAAACGGCGGCCGGCCGCTTATAAGTCTTCCTCACTTATTAACTTCTGTTC
CAGATCCTCTTCTGAGATGAGTTTTTGTTCGGGCCCCATGGAACCAAATAGTGAGCCACT
AAAAGCAAAGTTTTGAGCTGAACAATCTCTGAACTCTATATAAACTCTTCTTGGTTTAAC
ACTTAAATGATTGGAAAGGATTTTTGTAATTTTATCTGCTAGTAAAGAATTATTTGATCT
ATTTATTCCACCAATACTAGTTAATCTAACAAAGCAATATCCTTCATTACTTCCAGAAAA
TCGGAGGTTTTTTTGATAATCATAGTTGCTCATAATATATGCAACTGGCTTCCCTAAAAT
ATTAGATATAGCTAAAAAAAAAAATTTCATATATAAAATGTGTATATTTATAAATAGTAC
TAAAAGTAAATATACGAATATCATCATAAATAAGCTACATCCTATAACAGTTTTTTTCCA
CAACCATATTACGGTAATTATTTGATTTATATTTCTTACCATCTTCTATTTCGGATAGTG
TATTTTGAGCTTTATCGTCAGGGATAGAAATGTTTGTTATTAATTCACAGCACGGCATTT
TGGAAATTTTTTTTAAAAATATAAAGATTTTATATATTTATATTATTTAACTATAAAAAT
TGTAAATTTCTCTATTTTTATTTTATAAAAATATTTTATAAATTTGTAGTATGTTTTAAT
ACAAAGTATATACGTTAAAAAAATATATTTTATAAAAAAAATCGAAAAAAAAATTATAGG
ATTTTGATATTATATTTACAAATCAAATATATTATTTAGTTAAATAATTAAAGCACTAAT
TTTTCGAAAGTGCAATGAATTACTATTATTACTAAAAGGTTTGTATTTCTTAACACTCTT
GTGATATTAATCCATACACGCCATAATGCATTATTCCCACAATGTCAAAACACCTCGGGG
GATAAAAATATATTTTAAATATCAGCACATATTATATATATTATTGTGAATAAAACATCG
AAGCAAACAATGCCTTTTATACGTACTATAATATTATTTGTATTATTCAAATGGTGATAA
TTAAAAGTGGAATGAAATAAAAAAAAAAAATTTGAAGTTCAATATAATGAAGCAAAAAAA
TTAAAGTTTATTATAATATAGCTATATAAACATGAAAAATGAAAAAGAAAATGAAAATAA
AAAATAATAAAACAACAAATAACGAGTGAGCAATTTTAAATCATTTGTGAACGAATAGCA
TACGCAACACAAGCAACGTGTTTTTTGTAGTTTATCTTTTATTACATTTGGGTCTTATTT
TATATAGTATATTTTTATATATGAAAATATAAATATTTGGGGCATGCATTTATTTGATAT
AATTTAAAAAGGAGAGAGCCATAATGCCATTGTATTTGTAAAAATCTACTTAAAAAAAAA
TACAATATAAATAATTCCAAAGTATGTCTATCTCCTCATTTTCACCTTATATATATGATG
TGTGCAACTAAAAGCTTGGATTTGTATTTTTTTAATATTTAAAAAACTATACTGAAAAAA
AGCATAGGAAAAGGAGGTGTATGCATATATTTACATACATTTTATTAGCCTCAGAAAATT
AGAAAAAAAATGTATAATTATTTCAATTCATAATCCGTAATTTACTTATATATTATTTAT
AAAAAATTAATTATGAAATTCATATGTATTTTGAAGTGGATTATATGCACTATCATAAGA
AGCTGATTCGCTATAGTTTTGATTAAAGTCTTTTGGTATAAATGGTTGATAAGATTCGCG
GCCGCCACCGCGGTGGAGCTCCAATTCGCCCTATAGTGAGTCGTATTACGCGCGCTCACT
GGCCGTCGTTTTACAACGTCGTGACTGGGAAAACCCTGGCGTTACCCAACTTAATCGCCT
TGCAGCACATCCCCCTTTCGCCAGCTGGCGTAATAGCGAAGAGGCCCGCACCGATCGCCC
TTCCCAACAGTTGCGCAGCCTGAATGGCGAATGGGACGCGCCCTGTAGCGGCGCATTAAG
CGCGGCGGGTGTGGTGGTTACGCGCAGCGTGACCGCTACACTTGCCAGCGCCCTAGCGCC
CGCTCCTTTCGCTTTCTTCCCTTCCTTTCTCGCCACGTTCGCCGGCTTTCCCCGTCAAGC
TCTAAATCGGGGGCTCCCTTTAGGGTTCCGATTTAGTGCTTTACGGCACCTCGACCCCAA
AAAACTTGATTAGGGTGATGGTTCACGTAGTGGGCCATCGCCCTGATAGACGGTTTTTCG
CCCTTTGACGTTGGAGTCCACGTTCTTTAATAGTGGACTCTTGTTCCAAACTGGAACAAC
ACTCAACCCTATCTCGGTCTATTCTTTTGATTTATAAGGGATTTTGCCGATTTCGGCCTA
TTGGTTAAAAAATGAGCTGATTTAACAAAAATTTAACGCGAATTTTAACAAAATATTAAC
GCTTACAATTTAGGTGGCACTTTTCGGGGAAATGTGCGCGGAACCCCTATTTGTTTATTT
TTCTAAATACATTCAAATATGTATCCGCTCATGAGACAATAACCCTGATAAATGCTTCAA
TAATATTGAAAAAGGAAGAGTATGAGTATTCAACATTTCCGTGTCGCCCTTATTCCCTTT
TTTGCGGCATTTTGCCTTCCTGTTTTTGCTCACCCAGAAACGCTGGTGAAAGTAAAAGAT
GCTGAAGATCAGTTGGGTGCACGAGTGGGTTACATCGAACTGGATCTCAACAGCGGTAAG
ATCCTTGAGAGTTTTCGCCCCGAAGAACGTTTTCCAATGATGAGCACTTTTAAAGTTCTG
CTATGTGGCGCGGTATTATCCCGTATTGACGCCGGGCAAGAGCAACTCGGTCGCCGCATA
CACTATTCTCAGAATGACTTGGTTGAGTACTCACCAGTCACAGAAAAGCATCTTACGGAT
GGCATGACAGTAAGAGAATTATGCAGTGCTGCCATAACCATGAGTGATAACACTGCGGCC
AACTTACTTCTGACAACGATCGGAGGACCGAAGGAGCTAACCGCTTTTTTGCACAACATG
GGGGATCATGTAACTCGCCTTGATCGTTGGGAACCGGAGCTGAATGAAGCCATACCAAAC
GACGAGCGTGACACCACGATGCCTGTAGCAATGGCAACAACGTTGCGCAAACTATTAACT
GGCGAACTACTTACTCTAGCTTCCCGGCAACAATTAATAGACTGGATGGAGGCGGATAAA
GTTGCAGGACCACTTCTGCGCTCGGCCCTTCCGGCTGGCTGGTTTATTGCTGATAAATCT
GGAGCCGGTGAGCGTGGGTCTCGCGGTATCATTGCAGCACTGGGGCCAGATGGTAAGCCC
TCCCGTATCGTAGTTATCTACACGACGGGGAGTCAGGCAACTATGGATGAACGAAATAGA
CAGATCGCTGAGATAGGTGCCTCACTGATTAAGCATTGGTAACTGTCAGACCAAGTTTAC
TCATATATACTTTAGATTGATTTAAAACTTCATTTTTAATTTAAAAGGATCTAGGTGAAG
ATCCTTTTTGATAATCTCATGACCAAAATCCCTTAACGTGAGTTTTCGTTCCACTGAGCG
TCAGACCCCGTAGAAAAGATCAAAGGATCTTCTTGAGATCCTTTTTTTCTGCGCGTAATC
TGCTGCTTGCAAACAAAAAAACCACCGCTACCAGCGGTGGTTTGTTTGCCGGATCAAGAG
CTACCAACTCTTTTTCCGAAGGTAACTGGCTTCAGCAGAGCGCAGATACCAAATACTGTC
CTTCTAGTGTAGCCGTAGTTAGGCCACCACTTCAAGAACTCTGTAGCACCGCCTACATAC
CTCGCTCTGCTAATCCTGTTACCAGTGGCTGCTGCCAGTGGCGATAAGTCGTGTCTTACC
GGGTTGGACTCAAGACGATAGTTACCGGATAAGGCGCAGCGGTCGGGCTGAACGGGGGGT
TCGTGCACACAGCCCAGCTTGGAGCGAACGACCTACACCGAACTGAGATACCTACAGCGT
GAGCTATGAGAAAGCGCCACGCTTCCCGAAGGGAGAAAGGCGGACAGGTATCCGGTAAGC
GGCAGGGTCGGAACAGGAGAGCGCACGAGGGAGCTTCCAGGGGGAAACGCCTGGTATCTT
TATAGTCCTGTCGGGTTTCGCCACCTCTGACTTGAGCGTCGATTTTTGTGATGCTCGTCA
GGGGGGCGGAGCCTATGGAAAAACGCCAGCAACGCGGCCTTTTTACGGTTCCTGGCCTTT
TGCTGGCCTTTTGCTCACATGTTCTTTCCTGCGTTATCCCCTGATTCTGTGGATAACCGT
ATTACCGCCTTTGAGTGAGCTGATACCGCTCGCCGCAGCCGAACGACCGAGCGCAGCGAG
TCAGTGAGCGAGGAAGCGGAAGAGCGCCCAATACGCAAACCGCCTCTCCCCGCGCGTTGG
CCGATTCATTAATGCAGCTGGCACGACAGGTTTCCCGACTGGAAAGCGGGCAGTGAGCGC
AACGCAATTAATGTGAGTTAGCTCACTCATTAGGCACCCCAGGCTTTACACTTTATGCTT
CCGGCTCGTATGTTGTGTGGAATTGTGAGCGGATAACAATTTCACACAGGAAACAGCTAT
GACCATGATTACGCCAAGCTCGAAATTAACCCTCACTAAAGGGAACAAAAGCTGGTACC
Restriction sites to linearize plasmid SnaBI
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 1AAAGGTATTCCATGGAACCAAATAGTGAGCCACTAAAAGCA
Additional information primer 1GFP-tag MIF NcoI rev (in frame C-ter)
Sequence Primer 2TAAATCAAAGCGGCCGCGAATCTTATCAACCATTTATACC
Additional information primer 2GFP-tag MIF NotI fwd (1 kb upstream)
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