Summary

RMgm-709
Malaria parasiteP. berghei
Genotype
TaggedGene model (rodent): PBANKA_1200600; Gene model (P.falciparum): Not available; Gene product: Plasmodium exported protein, unknown function
Name tag: mCherry
Phenotype Asexual bloodstage;
Last modified: 2 December 2012, 15:34
  *RMgm-709
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
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-709
Principal name1955
Alternative name
Standardized name
Is the mutant parasite cloned after genetic modificationNo
Phenotype
Asexual blood stageThe tagged-protein shows a predominant location in the cytoplasm of the parasite
Gametocyte/GameteNot tested
Fertilization and ookineteNot tested
OocystNot tested
SporozoiteNot tested
Liver stageNot tested
Additional remarks phenotype

Mutant/mutation

Protein (function)

Phenotype

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.

Phenotype analyses of the blood stages by fluorescence microscopy show that the tagged-protein has a predominant location in the cytoplasm of the parasite.



Other mutants
RMgm-720: unsuccessful attempt to disrupt PBANKA_120060


  Tagged: Mutant parasite with a tagged gene
Details of the target gene
Gene Model of Rodent Parasite PBANKA_1200600
Gene Model P. falciparum ortholog Not available
Gene productPlasmodium exported protein, unknown function
Gene product: Alternative name
Details of the genetic modification
Name of the tagmCherry
Details of taggingC-terminal
Additional remarks: tagging
Commercial source of tag-antibodiesClontech
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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CTAAATTGTAAGCGTTAATATTTTGTTAAAATTCGCGTTAAATTTTTGTTAAATCAGCTC
ATTTTTTAACCAATAGGCCGAAATCGGCAAAATCCCTTATAAATCAAAAGAATAGACCGA
GATAGGGTTGAGTGTTGTTCCAGTTTGGAACAAGAGTCCACTATTAAAGAACGTGGACTC
CAACGTCAAAGGGCGAAAAACCGTCTATCAGGGCGATGGCCCACTACGTGAACCATCACC
CTAATCAAGTTTTTTGGGGTCGAGGTGCCGTAAAGCACTAAATCGGAACCCTAAAGGGAG
CCCCCGATTTAGAGCTTGACGGGGAAAGCCGGCGAACGTGGCGAGAAAGGAAGGGAAGAA
AGCGAAAGGAGCGGGCGCTAGGGCGCTGGCAAGTGTAGCGGTCACGCTGCGCGTAACCAC
CACACCCGCCGCGCTTAATGCGCCGCTACAGGGCGCGTCCCATTCGCCATTCAGGCTGCG
CAACTGTTGGGAAGGGCGATCGGTGCGGGCCTCTTCGCTATTACGCCAGCTGGCGAAAGG
GGGATGTGCTGCAAGGCGATTAAGTTGGGTAACGCCAGGGTTTTCCCAGTCACGACGTTG
TAAAACGACGGCCAGTGAGCGCGCGTAATACGACTCACTATAGGGCGAATTGGAGCTCCA
CCGCGGTGGCGGCCGCTCTAGAACTAGAGAATTATAAACTCTTACATTTTGCAATATCTA
TAATTATGTTTAGTTTATCAAAATTTTAATATACTCATTAATTAAATTAAATTTATTTGA
ATAAATTCATATTATATTATTTTATACTTTTGTGGACATTTACTAGTTTATGTTTTTTAT
ATACAACAATGAGCATCTTTTAAAATTAATATAAAATGATTATTAAAGAATTATATAAAT
ATCATATATTTTTCCATTTGAAAAACGTAATACCTTTATATATGTTTAATATTTAACCGA
TGTGTGCCAGAAGGAGAATGAAATCCGAGAATAGGATTAACAAAATCTTGATTATAAAAA
TTGGATTGTTAAACTTAATTTTCATATGGATATTAGGATATTTCACTAATGTAATATAAA
AATGCAAAATCTCAAAAATACGCAATAAGAAATTTAATATATTTGTGGTTATTTGCTCAT
TTGTTGTTTCTTTATTTCGATTAGTATTAATAAAACTATAATAATTTTCTTTTACCTTTT
TAGGACATCCCCGTTACATTGCGCAATGGTGAAATCGTAGTAAAAAATAAATTCAATTTA
CAGAGAATAAGAATATTAGCATCATGCTTTGAAGATGACACATACGATAGCAATAATAAT
AAAGATATACTTGCGAGAACTTATAATGATCCCATCTATTCATTTTATAAAACAAACCCT
AAAACTAAAGGACGTATTAATGAACGCAAAATATGCTTTATGAAAAAGATATATCCCAAT
ATTGATAAAGACGGTGTAAACTATGAAAGTCAATTTTTCGAACTAAGAAATAAAGTGCTG
GCTCCTATTGAAAAACGTAACCAAGGCCTACTAGAAGCTCATCATAATTTATTGAATTTT
GGTAATTATGTATCAGATATGAAATCTTATGGATTAGCAATAGAAGAAGTAAAGAAAAAT
ATTGATTCTTTTAGAAATACTTGTGAATCACAAATTTTAGGAATAAAAAATCTTTTATCA
AGTTATATGGATTCTTGTGGAACACAAATTGGATCCGGTGGAGGTGGAAGATCCATGGTG
AGCAAGGGCGAGGAGGATAACATGGCCATCATCAAGGAGTTCATGCGCTTCAAGGTGCAC
ATGGAGGGCTCCGTGAACGGCCACGAGTTCGAGATCGAGGGCGAGGGCGAGGGCCGCCCC
TACGAGGGCACCCAGACCGCCAAGCTGAAGGTGACCAAGGGTGGCCCCCTGCCCTTCGCC
TGGGACATCCTGTCCCCTCAGTTCATGTACGGCTCCAAGGCCTACGTGAAGCACCCCGCC
GACATCCCCGACTACTTGAAGCTGTCCTTCCCCGAGGGCTTCAAGTGGGAGCGCGTGATG
AACTTCGAGGACGGCGGCGTGGTGACCGTGACCCAGGACTCCTCCCTGCAGGACGGCGAG
TTCATCTACAAGGTGAAGCTGCGCGGCACCAACTTCCCCTCCGACGGCCCCGTAATGCAG
AAGAAGACCATGGGCTGGGAGGCCTCCTCCGAGCGGATGTACCCCGAGGACGGCGCCCTG
AAGGGCGAGATCAAGCAGAGGCTGAAGCTGAAGGACGGCGGCCACTACGACGCTGAGGTC
AAGACCACCTACAAGGCCAAGAAGCCCGTGCAGCTGCCCGGCGCCTACAACGTCAACATC
AAGTTGGACATCACCTCCCACAACGAGGACTACACCATCGTGGAACAGTACGAACGCGCC
GAGGGCCGCCACTCCACCGGCGGCATGGACGAGCTGTACAAGTAATCTAGAATTATTAAT
ATATATGAATATATATACATCGTTGTATGCGTAATATACGTGTAACTGTTTTTTTTAATT
TTTATATTAACTTTTCCTTATACATTTTAATGCCATTAACTAATAATATTTTACTTGAAA
AAATTCTAAATTTTTATAACTCTAATATAAAATCTGTGAATAATAAAAAATTAGCATAAA
GTACGAATTAAACATCGAATATTTGTTTTAAAGGGGCACACATTATGTGCATATAATTTT
ATTCTTTGAATTTAATTTTTTGCTTATAATTAGATTAAATTAATATAATTTTTTTATTGA
AAATAAAAACTTATTTTAAAAAATTAACAAATGGAAGTTAATGAAAAATACGAAAAAAGT
AAAGGGTTAATTCTTATATGGTCGGTACCGAGCTCGAATTCGATATCGAAATTGAAGGAA
AAAACATCATTTGTGTTTATATGTTTTTTTTTAATTTTTCAACTTTTTTTATTTAACTAT
GCTAATCATATTATAAACATTTTAAAAATCAAGTCCAACTATTTATGAATCATTGAAGAG
ACAACATAATTATAACAGTAAAAATAATAATAATAATAATAATTTTGTCTTATTGTTTCA
AACATATTATGTTCCATAAATATCTAAATTTTTAATATGGTCATAAATTCATAGGATTTA
ATTTATATAAAATATGGCTATTCATACTAGCCATTTTATGTGTGTAAAGTTTTTTTTTTC
TGAACAAAAAATAAAATGTTCATAATTTTAGCTATTTACATGCATGTGCATGCACAAAAA
AAAATATGCACACAACATACACATTTTTACAGTTATAAATACAATCAATTGGTATAAATA
TATAAGTAAGAAAAACGGGATCAAAGCTAGCTTCTGTATTTCCGCTAGACAGCCATCTCC
ATCTGGATTCGTCCGTGCGGGACGTAGCCCACGACCTCAAAATCCTCGGCGGTGAAATCG
TCGATTTCCTTGATGCGTTCCTTGTTGAGGATGTTCACAATGGGGAACGGTCTCGGTTCT
CTCCGCAGCTGCTCTTTTAAAGCCTCGACATGGTTCGTGTAGACATGCGTGTTCCCCATG
AAGTGAATGAACTCCTTAGGTTTTAGGTTGCAGACGTGTGCAACCATGAGCGTCAAAAGC
GAATAGGAAGCGATGTTGAAGGGGACGCCGAGGCCGACATCGCACGACCGCTGATACATG
ATGCACGACAGCTCCTTCTGGTCGTTCACGTAGAACTGGCACAACAAGTGACAAGGCGGC
AGCGCCATTTCGTCCAGCGCTGCAGGATTCCAGGCAGTCATGAGCATGCGACGATCTGTT
GGATTCGTTCTCAGCATCTGGATCACATTCTTCAGCTGGTCGACGCCCTGCCCTGTGTAG
TCTGTGTGCATGTCTTTGTATGCCGCGCCGAAGTGTCTCCACTGGAAGCCGTAGCCCGGG
CCGATGTCTCCGACCTCTCGGTGGGGGAGATTGCGCGAATCGAGGAACTCGCGTGTCACA
TTCTTGTCCCAGATCTTCACGCCCTTCTCAGAAAGATGGTTTGCGTTCGTGTCGCCGCGA
ATGAACCACAGCAACTCTTCGAGGACCCCTTTCCAGAACACACGCTTTGTGGTGAGAAGT
GGAAAGGCCTGATCCAGCGAGTAGCGCATAGTGCAGCCGAATTTGGAGATGACACCAACG
CCCGTTCGGTCATCCATTGTCCTTCCATTGTTAATAATGTCGGCAATGAGATCAAGGTAC
TGGAACTCTTCATGGCCTCTAAAGTGAACATGCGGAACGGCCCGAATCAGTTCCTTTTGC
TCGCGTTTTTTCCGGTCTTCTTCGTCCATCCACGCCAACACCGGGGCAATGGCTGCGGCC
GAAGAAGGAGCCTGCAACCCGTGCACGGGAGTTGTCTCCCTCGTGGACGTCAAGGAGCTC
ATTGCGTTGCTCGGTTCCGCAGTGGCTGCGTCGTCAGTCTTCCTTCTCTTCTCGAGAACC
ACAAAGTCGTAGGGAACCCCGTTGTCTGAGAAGGTCTTGGAAATGAAGATGGGTCGATAC
GTCGCTTCATTGTCCTTCTCTCTTCCGAGCTCCGGACAAAAGGGAACGAACACAGACTCG
GCAGGAGCTGCAGCCTGCGCAGCAGTTGATTTGTTTGAAAGAATGTCATCTCCGGGGAAC
GCAGGGAAGAAAACGTCGCACGGAAACTCGCGGGCTACACGCGTGATGTACAGGTGAGAG
GCAACGCCCAGAGACAGCGCTGCCTCGTACAGTCCCGCTCCTCCCACGACAAAAATCTGG
TCGACAGAATCCTTGTACTCTTCCTCCAGAAGGCTGAGAGCTGCTGGGAGTGAAGCACAG
ACTCGGACGCGCTGCTGGCCTTCAGCTTGAGGCTTCTCCGCCGCAATGTCTTCTTCTTTG
AGGGAAGAGGAAACGACGATGTTCAATCTGTCCACGAGGGGTCTAAACTTTCGAGGCATG
CTTTCCCAGGTTTTCCGTCCCATGACAACGGCGTTGAATCTCTTGCCGACTGATGGAGAG
GGAAGTCCAGAGTCGCCCGTCTTTGCAAATTTCCTGGGAAGCCACCCGTTCAGGCGACTG
GCTTCTTCGGGCGTCGTTTTTGTCACACGAGAAAAGTGTTTGAAATCTGTGGTCAAGTGG
GGCCACGGGAGGCCGTTGTTGATGCCGATGCCCCTCTTGGGGGTCATCGCGACGACCAGA
CACACCGGTTTATGCATTGATCCTTTGTAACATTTAGGTGTGTATTTATATATATAAGCA
ATATACAAATGAAAATACTATTATAAAACGGAAACTCAAAAAATGTTGAAACAAATAGTC
AATTTTTAAAATGAATGTGTAAATTGTTTAAGTTTTATGACAAAAAAAGGAAAAGTGTAT
ATAAAATAAAAATTGGAAAAAATTTATATATTACATGCCATATAATACCATTTATTTACT
ATTTATATCTTAATATAAAAAATTGTAGCAAAAATATATAATCGTATTTTATATATTTAA
TATTATATAACCCCTTAAAAAAAAAAATTTAATTGAAAAAATTTTAATTTATGCACTTTT
TTATTGTATTATATATATTCCGTTATTTTAATTACACATAAACTTAATAGAAAAATTAAT
TCGCTGCTAAATAAAAATTATAGGGCTATATATGATGGGATCATATTCTATCTTTCTCTT
TTTCAAATYCTTTATATTTTAGAATACAAATCGCTTTAAACAATAAAAAAGAGGTATATC
CTACCCGAATTGACACACACAATAAATAATAAATATATAAATAATAAAAATATAAATATA
TAAATTATAAATATATAAATTATATATATAAATATATTAACATGTGTGTAATTTGTTAAA
ATTGATATTAAACAGCTTTGCATGGGGAAAGAGTGGCTTTTATAAGCGAATCATATAATA
CAGTGTTAATTTTAATATATAAAATATAATTTCCTTAAATTGTTATTTAACATTTTTAAT
TGCCTACCCARGGTAATATAATATCCAAATGACAATCAAAGATAAATGAAATACCGCTCC
ATTTTTCCTATATATCCATTTTCATTTTGTGTAAATGTTCATAAAATTTTATCATATACT
CATAAAAAACAGGGGAAAAATGAAAAAATGAACGTCCACCCCGTGTGAATATGCTCATTT
TGTATACTATACATTTTTTTTTATATAATTTATATTTTCCAAATAAAAAAAATGTTTGTA
CATGTATATATCAATATTGTGTTATGGCATGATGAAAAACAATTATTTTATTCGTTTTTA
AATTTGTAGAATTTCCCTAAATTATTAATGATTTCATATTATAATATATGTATCTATATA
TTTTTAATGTGTAATTCAAAATGGCGAAAAAGAAACAAACCATACTTTATAAGCAACTCT
CTTTGAGCACTTACCATAAAAAATTATACTACTAAATTCAAATTAAATAAAAAAATATAT
ATTATAAAATTGGCTTTAATTATTTATAAGCACTTGCTTTGTTTTACTCAAAAAGTATTG
GCATAAGTATAAACATAAAACCAATTTTTCACAAAATATATTATATGTCCATACAACTAT
ATCCGAACYTTCAAAACACATCCAAATTTCCATTTACCACATGCAAGCAAAAGGATATTA
AATAAAAAAAAATTATTTATATTTTCATGATCACTTAAATAATATTTAAAAAAAAAAGAG
ACCATATATAAAATGTATAATTTAAATATAGCTAACTTATTAAAAAAAATGAATATTGAG
CATATATCCATAATTTAAAACACTTCTATTTTATGTTATTTGATATATAAGTCGATAAAT
TATTGCATGTATGCATATAGACAGATTAATATACACCAAAATAATAAAAATTATATATAG
TTCATATATTATTCTGAATAATAAATTAGCTTTCCTCTTAATTCACTTGGTATGTATATA
TATATATATATATATATATATATATATATGGATAAATAAATATAAGGTCTATAACAGGAA
AAAATGTGTATTAACTTTTTTTTATAATTATTTTATATTTTTTCTCCATTTATTTAAATA
TGTTAAAATTATTTCCCATTTTATAAACAAAAATTAATCCATAAAATAGCAAAATAACAA
AATGACAAAATAACAAAAACAAAAAAACGCATTATATGAGTTCATTTTACACAATCCAAA
GAAAAAGAAAATATGCTTAAAATATTCATAACACACTTTTAAGCTATAAAAATATATATA
TTTATTTGTGTCTATATTACCAACTCAATTTAATAGATGTGTTATGTGATTAATTCATAC
ACAAACATACAAAAATAAACACCATTTTTGAAAAGATTAATTTGAAATCATATATGTTAT
TATTATATATTTTTATATCAATGAAATTTAAAAAAATAAAAGGGAAATCAATGTATTAAA
AATAATTATATGTTATTTTATTTCCACAATATTTATTTATTATTTATTGTTGACCTGCAG
GCATGCAAGCTTATCGATACCGTCGACCTCGAGGGGGGGCCCGGTACCCAGCTTTTGTTC
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 1GCTCTAGAGAATTATAAACTCTTACATTTTGCAATATC
Additional information primer 1L6651 (XbaI); PBANKA_120060 Forward
Sequence Primer 2CGGGATCCAATTTGTGTTCCACAAGAATCCA
Additional information primer 2L6652 (BamHI); PBANKA_120060 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