Summary

RMgm-27
Malaria parasiteP. berghei
Genotype
TaggedGene model (rodent): PBANKA_1444000; Gene model (P.falciparum): PF3D7_1229400; Gene product: macrophage migration inhibitory factor (MIF)
Name tag: EGFP
Phenotype Asexual bloodstage; Gametocyte/Gamete; Fertilization and ookinete; Oocyst; Sporozoite;
Last modified: 10 April 2012, 19:31
  *RMgm-27
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-27
Principal name477
Alternative namepbMIF-GFP
Standardized name
Is the mutant parasite cloned after genetic modificationNo
Phenotype
Asexual blood stageExpression of MIF-GFP throughout asexual stages
Gametocyte/GameteNormal gametocytes; expression of MIF-GFP
Fertilization and ookineteNormal ookinetes; expression of MIF-GFP
OocystNormal oocysts; expression of MIF-GFP
SporozoiteNormal sporozoites; expression of MIF-GFP
Liver stageNot different from wild type
Additional remarks phenotype

Mutant/mutation
The mutant expresses a c-terminal GFP-tagged form of Macrophage Migration Inhibitory Factor protein.

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

Other mutants 
RMgm-31 is a transgenic mutant expressing c-myc-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 tagEGFP
Details of taggingC-terminal
Additional remarks: taggingA partial sequence of the 3'UTR of the P48/45 gene of P. berghei (PB001525.02.0) was used as the 3'UTR of the GFP-tagged gene.
Commercial source of tag-antibodiesRoche cat #: 11814460001
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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GGCCGCGAATCTTATCAACCATTTATACCAAAAGACTTTAATCAAAACTATAGCGAATCA
GCTTCTTATGATAGTGCATATAATCCACTTCAAAATACATATGAATTTCATAATTAATTT
TTTATAAATAATATATAAGTAAATTACGGATTATGAATTGAAATAATTATACATTTTTTT
TCTAATTTTCTGAGGCTAATAAAATGTATGTAAATATATGCATACACCTCCTTTTCCTAT
GCTTTTTTTCAGTATAGTTTTTTAAATATTAAAAAAATACAAATCCAAGCTTTTAGTTGC
ACACATCATATATATAAGGTGAAAATGAGGAGATAGACATACTTTGGAATTATTTATATT
GTATTTTTTTTTAAGTAGATTTTTACAAATACAATGGCATTATGGCTCTCTCCTTTTTAA
ATTATATCAAATAAATGCATGCCCCAAATATTTATATTTTCATATATAAAAATATACTAT
ATAAAATAAGACCCAAATGTAATAAAAGATAAACTACAAAAAACACGTTGCTTGTGTTGC
GTATGCTATTCGTTCACAAATGATTTAAAATTGCTCACTCGTTATTTGTTGTTTTATTAT
TTTTTATTTTCATTTTCTTTTTCATTTTTCATGTTTATATAGCTATATTATAATAAACTT
TAATTTTTTTGCTTCATTATATTGAACTTCAAATTTTTTTTTTTTATTTCATTCCACTTT
TAATTATCACCATTTGAATAATACAAATAATATTATAGTACGTATAAAAGGCATTGTTTG
CTTCGATGTTTTATTCACAATAATATATATAATATGTGCTGATATTTAAAATATATTTTT
ATCCCCCGAGGTGTTTTGACATTGTGGGAATAATGCATTATGGCGTGTATGGATTAATAT
CACAAGAGTGTTAAGAAATACAAACCTTTTAGTAATAATAGTAATTCATTGCACTTTCGA
AAAATTAGTGCTTTAATTATTTAACTAAATAATATATTTGATTTGTAAATATAATATCAA
AATCCTATAATTTTTTTTTCGATTTTTTTTATAAAATATATTTTTTTAACGTATATACTT
TGTATTAAAACATACTACAAATTTATAAAATATTTTTATAAAATAAAAATAGAGAAATTT
ACAATTTTTATAGTTAAATAATATAAATATATAAAATCTTTATATTTTTAAAAAAAATTT
CCAAAATGCCGTGCTGTGAATTAATAACAAACATTTCTATCCCTGACGATAAAGCTCAAA
ATACACTATCCGAAATAGAAGATGGTAAGAAATATAAATCAAATAATTACCGTAATATGG
TTGTGGAAAAAAACTGTTATAGGATGTAGCTTATTTATGATGATATTCGTATATTTACTT
TTAGTACTATTTATAAATATACACATTTTATATATGAAATTTTTTTTTTTAGCTATATCT
AATATTTTAGGGAAGCCAGTTGCATATATTATGAGCAACTATGATTATCAAAAAAACCTC
CGATTTTCTGGAAGTAATGAAGGATATTGCTTTGTTAGATTAACTAGTATTGGTGGAATA
AATAGATCAAATAATTCTTTACTAGCAGATAAAATTACAAAAATCCTTTCCAATCATTTA
AGTGTTAAACCAAGAAGAGTTTATATAGAGTTCAGAGATTGTTCAGCTCAAAACTTTGCT
TTTAGTGGCTCACTATTTGGTTCCATGGCTATGAGTAAAGGAGAAGAACTTTTCACTGGA
GTTGTCCCAATTCTTGTTGAATTAGATGGTGATGTTAATGGGCACAAATTTTCTGTCAGT
GGAGAGGGTGAAGGTGATGCAACATACGGAAAACTTACCCTTAAATTTATTTGCACTACT
GGAAAACTACCTGTTCCTTGGCCAACACTTGTCACTACTTTCGGTTATGGTGTTCAATGC
TTTGCGAGATACCCAGATCACATGAAACAGCATGACTTTTTCAAGAGTGCCATGCCCGAA
GGTTATGTACAGGAAAGAACTATATTTTTCAAAGATGACGGGAACTACAAGACACGTGCT
GAAGTCAAGTTTGAAGGTGATACCCTTGTTAATAGAATCGAGTTAAAAGGTATTGATTTT
AAAGAAGATGGAAACATTCTTGGACACAAATTGGAATACAACTATAACTCACACAATGTA
TACATCATGGCAGACAAACAAAAGAATGGAATCAAAGTTAACTTCAAAATTAGACACAAC
ATTGAAGATGGAAGCGTTCAACTAGCAGACCATTATCAACAAAATACTCCAATTGGCGAT
GGCCCTGTCCTTTTACCAGACAACCATTACCTGTCCACACAATCTGCCCTTTCGAAAGAT
CCCAACGAAAAGAGAGACCACATGGTCCTTCTTGAGTTTGTAACAGCTGCTGGGATTACA
CATGGCATGGATGAACTATACAAATAACATATGCTTAACATTCACATATATTAATAATTT
TAATTAATTTTTTTTTTTGCACAGATAAAACTTTTCTTTTTAAAAAAAAGGTATATACAC
TCTTTTTTACCCCACATTTAACTGATTTTCACTCACCATTACATATAGCTATATATTGCA
TATTACTATTATGTAAGGCAAATGTTTATTATTATTGCTGTTATTTTTTATTTATATATT
TTTTTCCATATATCAATTTAAGGAGAAATAAAATAATTTGCTATTTGAAAAAATATAAAA
CAAATATCTAGTCGTGCAATACTGAATTCGAATACTTAAAACAAAAAATAAATTAATTAA
ATACCCTTTTAATATTGTAGTTAAAATCGATGTGTTTAGAGAATATTTTTTATTGCTATT
TATCTTCTTTTTTTTATTTTTCTTCCATGTTTTCTCATTCAGCTTTCTATGTATTTTAGT
AATATTCTCCACACACAAACTATAAATTTTCTTATGAAAAATCACAAAATTTAGCAAATA
GCGCTTAGGCATGTACTCGTAAATAAGCATGTGCATAGTATAATACACTTCTCATCAATT
TTGAGAGGATTTTATTTAATTTCGGCACGCCCCTCTCAACTTGTGTTTATCACATATTCA
ATACTCCATAAAAAATGATAGCATCATTTCGATATGCGGATATCGAAATTGAAGGAAAAA
ACATCATTTGTGTTTATATGTTTTTTTTTAATTTTTCAACTTTTTTTATTTAACTATGCT
AATCATATTATAAACATTTTAAAAATCAAGTCCAACTATTTATGAATCATTGAAGAGACA
ACATAATTATAACAGTAAAAATAATAATAATAATAATAATTTTGTCTTATTGTTTCAAAC
ATATTATGTTCCATAAATATCTAAATTTTTAATATGGTCATAAATTCATAGGATTTAATT
TATATAAAATATGGCTATTCATACTAGCCATTTTATGTGTGTAAAGTTTTTTTTTTCTGA
ACAAAAAATAAAATGTTCATAATTTTAGCTATTTACATGCATGTGCATGCACAAAAAAAA
ATATGCACACAACATACACATTTTTACAGTTATAAATACAATCAATTGGTATAAATATAT
AAGTAAGAAAAACGGGATCAAAGCTAGCTTCTGTATTTCCGCTAGACAGCCATCTCCATC
TGGATTCGTCCGTGCGGGACGTAGCCCACGACCTCAAAATCCTCGGCGGTGAAATCGTCG
ATTTCCTTGATGCGTTCCTTGTTGAGGATGTTCACAATGGGGAACGGTCTCGGTTCTCTC
CGCAGCTGCTCTTTTAAAGCCTCGACATGGTTCGTGTAGACATGCGTGTTCCCCATGAAG
TGAATGAACTCCTTAGGTTTTAGGTTGCAGACGTGTGCAACCATGAGCGTCAAAAGCGAA
TAGGAAGCGATGTTGAAGGGGACGCCGAGGCCGACATCGCACGACCGCTGATACATGATG
CACGACAGCTCCTTCTGGTCGTTCACGTAGAACTGGCACAACAAGTGACAAGGCGGCAGC
GCCATTTCGTCCAGCGCTGCAGGATTCCAGGCAGTCATGAGCATGCGACGATCTGTTGGA
TTCGTTCTCAGCATCTGGATCACATTCTTCAGCTGGTCGACGCCCTGCCCTGTGTAGTCT
GTGTGCATGTCTTTGTATGCCGCGCCGAAGTGTCTCCACTGGAAGCCGTAGCCCGGGCCG
ATGTCTCCGACCTCTCGGTGGGGGAGATTGCGCGAATCGAGGAACTCGCGTGTCACATTC
TTGTCCCAGATCTTCACGCCCTTCTCAGAAAGATGGTTTGCGTTCGTGTCGCCGCGAATG
AACCACAGCAACTCTTCGAGGACCCCTTTCCAGAACACACGCTTTGTGGTGAGAAGTGGA
AAGGCCTGATCCAGCGAGTAGCGCATAGTGCAGCCGAATTTGGAGATGACACCAACGCCC
GTTCGGTCATCCATTGTCCTTCCATTGTTAATAATGTCGGCAATGAGATCAAGGTACTGG
AACTCTTCATGGCCTCTAAAGTGAACATGCGGAACGGCCCGAATCAGTTCCTTTTGCTCG
CGTTTTTTCCGGTCTTCTTCGTCCATCCACGCCAACACCGGGGCAATGGCTGCGGCCGAA
GAAGGAGCCTGCAACCCGTGCACGGGAGTTGTCTCCCTCGTGGACGTCAAGGAGCTCATT
GCGTTGCTCGGTTCCGCAGTGGCTGCGTCGTCAGTCTTCCTTCTCTTCTCGAGAACCACA
AAGTCGTAGGGAACCCCGTTGTCTGAGAAGGTCTTGGAAATGAAGATGGGTCGATACGTC
GCTTCATTGTCCTTCTCTCTTCCGAGCTCCGGACAAAAGGGAACGAACACAGACTCGGCA
GGAGCTGCAGCCTGCGCAGCAGTTGATTTGTTTGAAAGAATGTCATCTCCGGGGAACGCA
GGGAAGAAAACGTCGCACGGAAACTCGCGGGCTACACGCGTGATGTACAGGTGAGAGGCA
ACGCCCAGAGACAGCGCTGCCTCGTACAGTCCCGCTCCTCCCACGACAAAAATCTGGTCG
ACAGAATCCTTGTACTCTTCCTCCAGAAGGCTGAGAGCTGCTGGGAGTGAAGCACAGACT
CGGACGCGCTGCTGGCCTTCAGCTTGAGGCTTCTCCGCCGCAATGTCTTCTTCTTTGAGG
GAAGAGGAAACGACGATGTTCAATCTGTCCACGAGGGGTCTAAACTTTCGAGGCATGCTT
TCCCAGGTTTTCCGTCCCATGACAACGGCGTTGAATCTCTTGCCGACTGATGGAGAGGGA
AGTCCAGAGTCGCCCGTCTTTGCAAATTTCCTGGGAAGCCACCCGTTCAGGCGACTGGCT
TCTTCGGGCGTCGTTTTTGTCACACGAGAAAAGTGTTTGAAATCTGTGGTCAAGTGGGGC
CACGGGAGGCCGTTGTTGATGCCGATGCCCCTCTTGGGGGTCATCGCGACGACCAGACAC
ACCGGTTTATGCATTGATCCTTTGTAACATTTAGGTGTGTATTTATATATATAAGCAATA
TACAAATGAAAATACTATTATAAAACGGAAACTCAAAAAATGTTGAAACAAATAGTCAAT
TTTTAAAATGAATGTGTAAATTGTTTAAGTTTTATGACAAAAAAAGGAAAAGTGTATATA
AAATAAAAATTGGAAAAAATTTATATATTACATGCCATATAATACCATTTATTTACTATT
TATATCTTAATATAAAAAATTGTAGCAAAAATATATAATCGTATTTTATATATTTAATAT
TATATAACCCCTTAAAAAAAAAAATTTAATTGAAAAAATTTTAATTTATGCACTTTTTTA
TTGTATTATATATATTCCGTTATTTTAATTACACATAAACTTAATAGAAAAATTAATTCG
CTGCTAAATAAAAATTATAGGGCTATATATGATGGGATCATATTCTATCTTTCTCTTTTT
CAAATYCTTTATATTTTAGAATACAAATCGCTTTAAACAATAAAAAAGAGGTATATCCTA
CCCGAATTGACACACACAATAAATAATAAATATATAAATAATAAAAATATAAATATATAA
ATTATAAATATATAAATTATATATATAAATATATTAACATGTGTGTAATTTGTTAAAATT
GATATTAAACAGCTTTGCATGGGGAAAGAGTGGCTTTTATAAGCGAATCATATAATACAG
TGTTAATTTTAATATATAAAATATAATTTCCTTAAATTGTTATTTAACATTTTTAATTGC
CTACCCARGGTAATATAATATCCAAATGACAATCAAAGATAAATGAAATACCGCTCCATT
TTTCCTATATATCCATTTTCATTTTGTGTAAATGTTCATAAAATTTTATCATATACTCAT
AAAAAACAGGGGAAAAATGAAAAAATGAACGTCCACCCCGTGTGAATATGCTCATTTTGT
ATACTATACATTTTTTTTTATATAATTTATATTTTCCAAATAAAAAAAATGTTTGTACAT
GTATATATCAATATTGTGTTATGGCATGATGAAAAACAATTATTTTATTCGTTTTTAAAT
TTGTAGAATTTCCCTAAATTATTAATGATTTCATATTATAATATATGTATCTATATATTT
TTAATGTGTAATTCAAAATGGCGAAAAAGAAACAAACCATACTTTATAAGCAACTCTCTT
TGAGCACTTACCATAAAAAATTATACTACTAAATTCAAATTAAATAAAAAAATATATATT
ATAAAATTGGCTTTAATTATTTATAAGCACTTGCTTTGTTTTACTCAAAAAGTATTGGCA
TAAGTATAAACATAAAACCAATTTTTCACAAAATATATTATATGTCCATACAACTATATC
CGAACYTTCAAAACACATCCAAATTTCCATTTACCACATGCAAGCAAAAGGATATTAAAT
AAAAAAAAATTATTTATATTTTCATGATCACTTAAATAATATTTAAAAAAAAAAGAGACC
ATATATAAAATGTATAATTTAAATATAGCTAACTTATTAAAAAAAATGAATATTGAGCAT
ATATCCATAATTTAAAACACTTCTATTTTATGTTATTTGATATATAAGTCGATAAATTAT
TGCATGTATGCATATAGACAGATTAATATACACCAAAATAATAAAAATTATATATAGTTC
ATATATTATTCTGAATAATAAATTAGCTTTCCTCTTAATTCACTTGGTATGTATATATAT
ATATATATATATATATATATATATATGGATAAATAAATATAAGGTCTATAACAGGAAAAA
ATGTGTATTAACTTTTTTTTATAATTATTTTATATTTTTTCTCCATTTATTTAAATATGT
TAAAATTATTTCCCATTTTATAAACAAAAATTAATCCATAAAATAGCAAAATAACAAAAT
GACAAAATAACAAAAACAAAAAAACGCATTATATGAGTTCATTTTACACAATCCAAAGAA
AAAGAAAATATGCTTAAAATATTCATAACACACTTTTAAGCTATAAAAATATATATATTT
ATTTGTGTCTATATTACCAACTCAATTTAATAGATGTGTTATGTGATTAATTCATACACA
AACATACAAAAATAAACACCATTTTTGAAAAGATTAATTTGAAATCATATATGTTATTAT
TATATATTTTTATATCAATGAAATTTAAAAAAATAAAAGGGAAATCAATGTATTAAAAAT
AATTATATGTTATTTTATTTCCACAATATTTATTTATTATTTATTGTTGACCTGCAGGCA
TGCAAGCTTATCGATACCGTCGACCTCGAGGGGGGGCCCGGTACCAGCTTTTGTTCCCTT
TAGTGAGGGTTAATTTCGAGCTTGGCGTAATCATGGTCATAGCTGTTTCCTGTGTGAAAT
TGTTATCCGCTCACAATTCCACACAACATACGAGCCGGAAGCATAAAGTGTAAAGCCTGG
GGTGCCTAATGAGTGAGCTAACTCACATTAATTGCGTTGCGCTCACTGCCCGCTTTCCAG
TCGGGAAACCTGTCGTGCCAGCTGCATTAATGAATCGGCCAACGCGCGGGGAGAGGCGGT
TTGCGTATTGGGCGCTCTTCCGCTTCCTCGCTCACTGACTCGCTGCGCTCGGTCGTTCGG
CTGCGGCGAGCGGTATCAGCTCACTCAAAGGCGGTAATACGGTTATCCACAGAATCAGGG
GATAACGCAGGAAAGAACATGTGAGCAAAAGGCCAGCAAAAGGCCAGGAACCGTAAAAAG
GCCGCGTTGCTGGCGTTTTTCCATAGGCTCCGCCCCCCTGACGAGCATCACAAAAATCGA
CGCTCAAGTCAGAGGTGGCGAAACCCGACAGGACTATAAAGATACCAGGCGTTTCCCCCT
GGAAGCTCCCTCGTGCGCTCTCCTGTTCCGACCCTGCCGCTTACCGGATACCTGTCCGCC
TTTCTCCCTTCGGGAAGCGTGGCGCTTTCTCATAGCTCACGCTGTAGGTATCTCAGTTCG
GTGTAGGTCGTTCGCTCCAAGCTGGGCTGTGTGCACGAACCCCCCGTTCAGCCCGACCGC
TGCGCCTTATCCGGTAACTATCGTCTTGAGTCCAACCCGGTAAGACACGACTTATCGCCA
CTGGCAGCAGCCACTGGTAACAGGATTAGCAGAGCGAGGTATGTAGGCGGTGCTACAGAG
TTCTTGAAGTGGTGGCCTAACTACGGCTACACTAGAAGGACAGTATTTGGTATCTGCGCT
CTGCTGAAGCCAGTTACCTTCGGAAAAAGAGTTGGTAGCTCTTGATCCGGCAAACAAACC
ACCGCTGGTAGCGGTGGTTTTTTTGTTTGCAAGCAGCAGATTACGCGCAGAAAAAAAGGA
TCTCAAGAAGATCCTTTGATCTTTTCTACGGGGTCTGACGCTCAGTGGAACGAAAACTCA
CGTTAAGGGATTTTGGTCATGAGATTATCAAAAAGGATCTTCACCTAGATCCTTTTAAAT
TAAAAATGAAGTTTTAAATCAATCTAAAGTATATATGAGTAAACTTGGTCTGACAGTTAC
CAATGCTTAATCAGTGAGGCACCTATCTCAGCGATCTGTCTATTTCGTTCATCCATAGTT
GCCTGACTCCCCGTCGTGTAGATAACTACGATACGGGAGGGCTTACCATCTGGCCCCAGT
GCTGCAATGATACCGCGAGACCCACGCTCACCGGCTCCAGATTTATCAGCAATAAACCAG
CCAGCCGGAAGGGCCGAGCGCAGAAGTGGTCCTGCAACTTTATCCGCCTCCATCCAGTCT
ATTAATTGTTGCCGGGAAGCTAGAGTAAGTAGTTCGCCAGTTAATAGTTTGCGCAACGTT
GTTGCCATTGCTACAGGCATCGTGGTGTCACGCTCGTCGTTTGGTATGGCTTCATTCAGC
TCCGGTTCCCAACGATCAAGGCGAGTTACATGATCCCCCATGTTGTGCAAAAAAGCGGTT
AGCTCCTTCGGTCCTCCGATCGTTGTCAGAAGTAAGTTGGCCGCAGTGTTATCACTCATG
GTTATGGCAGCACTGCATAATTCTCTTACTGTCATGCCATCCGTAAGATGCTTTTCTGTG
ACTGGTGAGTACTCAACCAAGTCATTCTGAGAATAGTGTATGCGGCGACCGAGTTGCTCT
TGCCCGGCGTCAATACGGGATAATACCGCGCCACATAGCAGAACTTTAAAAGTGCTCATC
ATTGGAAAACGTTCTTCGGGGCGAAAACTCTCAAGGATCTTACCGCTGTTGAGATCCAGT
TCGATGTAACCCACTCGTGCACCCAACTGATCTTCAGCATCTTTTACTTTCACCAGCGTT
TCTGGGTGAGCAAAAACAGGAAGGCAAAATGCCGCAAAAAAGGGAATAAGGGCGACACGG
AAATGTTGAATACTCATACTCTTCCTTTTTCAATATTATTGAAGCATTTATCAGGGTTAT
TGTCTCATGAGCGGATACATATTTGAATGTATTTAGAAAAATAAACAAATAGGGGTTCCG
CGCACATTTCCCCGAAAAGTGCCACCTAAATTGTAAGCGTTAATATTTTGTTAAAATTCG
CGTTAAATTTTTGTTAAATCAGCTCATTTTTTAACCAATAGGCCGAAATCGGCAAAATCC
CTTATAAATCAAAAGAATAGACCGAGATAGGGTTGAGTGTTGTTCCAGTTTGGAACAAGA
GTCCACTATTAAAGAACGTGGACTCCAACGTCAAAGGGCGAAAAACCGTCTATCAGGGCG
ATGGCCCACTACGTGAACCATCACCCTAATCAAGTTTTTTGGGGTCGAGGTGCCGTAAAG
CACTAAATCGGAACCCTAAAGGGAGCCCCCGATTTAGAGCTTGACGGGGAAAGCCGGCGA
ACGTGGCGAGAAAGGAAGGGAAGAAAGCGAAAGGAGCGGGCGCTAGGGCGCTGGCAAGTG
TAGCGGTCACGCTGCGCGTAACCACCACACCCGCCGCGCTTAATGCGCCGCTACAGGGCG
CGTCCCATTCGCCATTCAGGCTGCGCAACTGTTGGGAAGGGCGATCGGTGCGGGCCTCTT
CGCTATTACGCCAGCTGGCGAAAGGGGGATGTGCTGCAAGGCGATTAAGTTGGGTAACGC
CAGGGTTTTCCCAGTCACGACGTTGTAAAACGACGGCCAGTGAGCGCGCGTAATACGACT
CACTATAGGGCGAATTGGAGCTCCACCGCGGTGGC
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 modificationA partial sequence of the 3'UTR of the P48/45 gene of P. berghei (PB001525.02.0) was used as the 3'UTR of the GFP-tagged gene.
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