Summary

RMgm-710
Malaria parasiteP. berghei
Genotype
TaggedGene model (rodent): PBANKA_1137800; Gene model (P.falciparum): PF3D7_1361800; Gene product: glideosome-associated connector
Name tag: mCherry
Phenotype Asexual bloodstage;
Last modified: 2 December 2012, 15:40
  *RMgm-710
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-710
Principal name1919
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-725: mutant lacking expression of PBANKA_113780


  Tagged: Mutant parasite with a tagged gene
Details of the target gene
Gene Model of Rodent Parasite PBANKA_1137800
Gene Model P. falciparum ortholog PF3D7_1361800
Gene productglideosome-associated connector
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
CCGCGGTGGCGGCCGCTCTAGAACTAGTTGTGGTGGAATACCTACTATAGCAAATTTGAT
ATCTAAATCTGTTAAAAAGTCAATTGTAGTTTTAGAAGCATTACGTATATTATTATGTGT
ATTATATTATACAAAAAATGTCGAAGGTGTAGTTAATGAATATTCTGAAGAAGACGAAGA
ATTATTCAATGCTAGATTAGGGGGATGGTATAATATAAGTATGGACAAAGAAATGATAGA
TGTAATATTACAATCTGTTTTAACATGTTCAAATGATGAGAATCATCAAAAACAATTGAG
ATTACAAAAAGTGTCTCTTGGTTTATTAGCTTATTTTGCATATCATAGGTTAGGTATTAT
TTCTATGACTGCATCTGGTTTTGATATTTTGGCTAAAAACAATTTAAATCATTTCGGTGG
TGATATGGTAATTATGCAATTGTTGGCTATATGTATAGATAACATTGCGATGAATTCAGC
AGAAATATACGATATGACAATTACGAGAGATATTATAAAATTATTTAAATCATCTATATC
TAAAATCCCAAATAAAAAAGATAATAAGCAAATTGTACAAAGTATTGAAAAAACAGTAGA
AGCAATGGGTTCAGATGGAGATCCTTTAGATGCATTTAAAGATACTATATTAACTTTTGA
TTTTAGTTTATCAGAATTTGATAAAGATCCATATGTTAATGGTGTACACGATTTACCTCA
AAATATCAAAGAAGCGTTGAGAACAGGTGGTCAATATAAAATATACCATAAAAGTGATAA
AAGAACATTGTTCAAATGGAAAGCTAGTCAAGATTTAGGGACCTTAGAATGGACAGTAGG
AGAAAATGTAGATCGTATATTTAAAATATCAGTAGTACGTATTAAAAATATATCCAAAGG
ATTAGTACATCCTTTGTTAAAAGCGGCTAATAAATATGAGCCTAGAAAAGTCAATTCAAA
AGTAGTATTATGCGTTTATGGACCCCCTACTGAAGATTTTCCAGAGGGCCTAGAGTTGCC
AATTAAAACAAAAACAAACAAAGAAAGAGATGCATTCGCCGATCTATTAATTCTTTGGCG
TGATGCTGCTAGTTATAACTATGGATCCGGTGGAGGTGGAAGATCCATGGTGAGCAAGGG
CGAGGAGGATAACATGGCCATCATCAAGGAGTTCATGCGCTTCAAGGTGCACATGGAGGG
CTCCGTGAACGGCCACGAGTTCGAGATCGAGGGCGAGGGCGAGGGCCGCCCCTACGAGGG
CACCCAGACCGCCAAGCTGAAGGTGACCAAGGGTGGCCCCCTGCCCTTCGCCTGGGACAT
CCTGTCCCCTCAGTTCATGTACGGCTCCAAGGCCTACGTGAAGCACCCCGCCGACATCCC
CGACTACTTGAAGCTGTCCTTCCCCGAGGGCTTCAAGTGGGAGCGCGTGATGAACTTCGA
GGACGGCGGCGTGGTGACCGTGACCCAGGACTCCTCCCTGCAGGACGGCGAGTTCATCTA
CAAGGTGAAGCTGCGCGGCACCAACTTCCCCTCCGACGGCCCCGTAATGCAGAAGAAGAC
CATGGGCTGGGAGGCCTCCTCCGAGCGGATGTACCCCGAGGACGGCGCCCTGAAGGGCGA
GATCAAGCAGAGGCTGAAGCTGAAGGACGGCGGCCACTACGACGCTGAGGTCAAGACCAC
CTACAAGGCCAAGAAGCCCGTGCAGCTGCCCGGCGCCTACAACGTCAACATCAAGTTGGA
CATCACCTCCCACAACGAGGACTACACCATCGTGGAACAGTACGAACGCGCCGAGGGCCG
CCACTCCACCGGCGGCATGGACGAGCTGTACAAGTAATCTAGAATTATTAATATATATGA
ATATATATACATCGTTGTATGCGTAATATACGTGTAACTGTTTTTTTTAATTTTTATATT
AACTTTTCCTTATACATTTTAATGCCATTAACTAATAATATTTTACTTGAAAAAATTCTA
AATTTTTATAACTCTAATATAAAATCTGTGAATAATAAAAAATTAGCATAAAGTACGAAT
TAAACATCGAATATTTGTTTTAAAGGGGCACACATTATGTGCATATAATTTTATTCTTTG
AATTTAATTTTTTGCTTATAATTAGATTAAATTAATATAATTTTTTTATTGAAAATAAAA
ACTTATTTTAAAAAATTAACAAATGGAAGTTAATGAAAAATACGAAAAAAGTAAAGGGTT
AATTCTTATATGGTCGGTACCGAGCTCGAATTCGATATCGAAATTGAAGGAAAAAACATC
ATTTGTGTTTATATGTTTTTTTTTAATTTTTCAACTTTTTTTATTTAACTATGCTAATCA
TATTATAAACATTTTAAAAATCAAGTCCAACTATTTATGAATCATTGAAGAGACAACATA
ATTATAACAGTAAAAATAATAATAATAATAATAATTTTGTCTTATTGTTTCAAACATATT
ATGTTCCATAAATATCTAAATTTTTAATATGGTCATAAATTCATAGGATTTAATTTATAT
AAAATATGGCTATTCATACTAGCCATTTTATGTGTGTAAAGTTTTTTTTTTCTGAACAAA
AAATAAAATGTTCATAATTTTAGCTATTTACATGCATGTGCATGCACAAAAAAAAATATG
CACACAACATACACATTTTTACAGTTATAAATACAATCAATTGGTATAAATATATAAGTA
AGAAAAACGGGATCAAAGCTAGCTTCTGTATTTCCGCTAGACAGCCATCTCCATCTGGAT
TCGTCCGTGCGGGACGTAGCCCACGACCTCAAAATCCTCGGCGGTGAAATCGTCGATTTC
CTTGATGCGTTCCTTGTTGAGGATGTTCACAATGGGGAACGGTCTCGGTTCTCTCCGCAG
CTGCTCTTTTAAAGCCTCGACATGGTTCGTGTAGACATGCGTGTTCCCCATGAAGTGAAT
GAACTCCTTAGGTTTTAGGTTGCAGACGTGTGCAACCATGAGCGTCAAAAGCGAATAGGA
AGCGATGTTGAAGGGGACGCCGAGGCCGACATCGCACGACCGCTGATACATGATGCACGA
CAGCTCCTTCTGGTCGTTCACGTAGAACTGGCACAACAAGTGACAAGGCGGCAGCGCCAT
TTCGTCCAGCGCTGCAGGATTCCAGGCAGTCATGAGCATGCGACGATCTGTTGGATTCGT
TCTCAGCATCTGGATCACATTCTTCAGCTGGTCGACGCCCTGCCCTGTGTAGTCTGTGTG
CATGTCTTTGTATGCCGCGCCGAAGTGTCTCCACTGGAAGCCGTAGCCCGGGCCGATGTC
TCCGACCTCTCGGTGGGGGAGATTGCGCGAATCGAGGAACTCGCGTGTCACATTCTTGTC
CCAGATCTTCACGCCCTTCTCAGAAAGATGGTTTGCGTTCGTGTCGCCGCGAATGAACCA
CAGCAACTCTTCGAGGACCCCTTTCCAGAACACACGCTTTGTGGTGAGAAGTGGAAAGGC
CTGATCCAGCGAGTAGCGCATAGTGCAGCCGAATTTGGAGATGACACCAACGCCCGTTCG
GTCATCCATTGTCCTTCCATTGTTAATAATGTCGGCAATGAGATCAAGGTACTGGAACTC
TTCATGGCCTCTAAAGTGAACATGCGGAACGGCCCGAATCAGTTCCTTTTGCTCGCGTTT
TTTCCGGTCTTCTTCGTCCATCCACGCCAACACCGGGGCAATGGCTGCGGCCGAAGAAGG
AGCCTGCAACCCGTGCACGGGAGTTGTCTCCCTCGTGGACGTCAAGGAGCTCATTGCGTT
GCTCGGTTCCGCAGTGGCTGCGTCGTCAGTCTTCCTTCTCTTCTCGAGAACCACAAAGTC
GTAGGGAACCCCGTTGTCTGAGAAGGTCTTGGAAATGAAGATGGGTCGATACGTCGCTTC
ATTGTCCTTCTCTCTTCCGAGCTCCGGACAAAAGGGAACGAACACAGACTCGGCAGGAGC
TGCAGCCTGCGCAGCAGTTGATTTGTTTGAAAGAATGTCATCTCCGGGGAACGCAGGGAA
GAAAACGTCGCACGGAAACTCGCGGGCTACACGCGTGATGTACAGGTGAGAGGCAACGCC
CAGAGACAGCGCTGCCTCGTACAGTCCCGCTCCTCCCACGACAAAAATCTGGTCGACAGA
ATCCTTGTACTCTTCCTCCAGAAGGCTGAGAGCTGCTGGGAGTGAAGCACAGACTCGGAC
GCGCTGCTGGCCTTCAGCTTGAGGCTTCTCCGCCGCAATGTCTTCTTCTTTGAGGGAAGA
GGAAACGACGATGTTCAATCTGTCCACGAGGGGTCTAAACTTTCGAGGCATGCTTTCCCA
GGTTTTCCGTCCCATGACAACGGCGTTGAATCTCTTGCCGACTGATGGAGAGGGAAGTCC
AGAGTCGCCCGTCTTTGCAAATTTCCTGGGAAGCCACCCGTTCAGGCGACTGGCTTCTTC
GGGCGTCGTTTTTGTCACACGAGAAAAGTGTTTGAAATCTGTGGTCAAGTGGGGCCACGG
GAGGCCGTTGTTGATGCCGATGCCCCTCTTGGGGGTCATCGCGACGACCAGACACACCGG
TTTATGCATTGATCCTTTGTAACATTTAGGTGTGTATTTATATATATAAGCAATATACAA
ATGAAAATACTATTATAAAACGGAAACTCAAAAAATGTTGAAACAAATAGTCAATTTTTA
AAATGAATGTGTAAATTGTTTAAGTTTTATGACAAAAAAAGGAAAAGTGTATATAAAATA
AAAATTGGAAAAAATTTATATATTACATGCCATATAATACCATTTATTTACTATTTATAT
CTTAATATAAAAAATTGTAGCAAAAATATATAATCGTATTTTATATATTTAATATTATAT
AACCCCTTAAAAAAAAAAATTTAATTGAAAAAATTTTAATTTATGCACTTTTTTATTGTA
TTATATATATTCCGTTATTTTAATTACACATAAACTTAATAGAAAAATTAATTCGCTGCT
AAATAAAAATTATAGGGCTATATATGATGGGATCATATTCTATCTTTCTCTTTTTCAAAT
YCTTTATATTTTAGAATACAAATCGCTTTAAACAATAAAAAAGAGGTATATCCTACCCGA
ATTGACACACACAATAAATAATAAATATATAAATAATAAAAATATAAATATATAAATTAT
AAATATATAAATTATATATATAAATATATTAACATGTGTGTAATTTGTTAAAATTGATAT
TAAACAGCTTTGCATGGGGAAAGAGTGGCTTTTATAAGCGAATCATATAATACAGTGTTA
ATTTTAATATATAAAATATAATTTCCTTAAATTGTTATTTAACATTTTTAATTGCCTACC
CARGGTAATATAATATCCAAATGACAATCAAAGATAAATGAAATACCGCTCCATTTTTCC
TATATATCCATTTTCATTTTGTGTAAATGTTCATAAAATTTTATCATATACTCATAAAAA
ACAGGGGAAAAATGAAAAAATGAACGTCCACCCCGTGTGAATATGCTCATTTTGTATACT
ATACATTTTTTTTTATATAATTTATATTTTCCAAATAAAAAAAATGTTTGTACATGTATA
TATCAATATTGTGTTATGGCATGATGAAAAACAATTATTTTATTCGTTTTTAAATTTGTA
GAATTTCCCTAAATTATTAATGATTTCATATTATAATATATGTATCTATATATTTTTAAT
GTGTAATTCAAAATGGCGAAAAAGAAACAAACCATACTTTATAAGCAACTCTCTTTGAGC
ACTTACCATAAAAAATTATACTACTAAATTCAAATTAAATAAAAAAATATATATTATAAA
ATTGGCTTTAATTATTTATAAGCACTTGCTTTGTTTTACTCAAAAAGTATTGGCATAAGT
ATAAACATAAAACCAATTTTTCACAAAATATATTATATGTCCATACAACTATATCCGAAC
YTTCAAAACACATCCAAATTTCCATTTACCACATGCAAGCAAAAGGATATTAAATAAAAA
AAAATTATTTATATTTTCATGATCACTTAAATAATATTTAAAAAAAAAAGAGACCATATA
TAAAATGTATAATTTAAATATAGCTAACTTATTAAAAAAAATGAATATTGAGCATATATC
CATAATTTAAAACACTTCTATTTTATGTTATTTGATATATAAGTCGATAAATTATTGCAT
GTATGCATATAGACAGATTAATATACACCAAAATAATAAAAATTATATATAGTTCATATA
TTATTCTGAATAATAAATTAGCTTTCCTCTTAATTCACTTGGTATGTATATATATATATA
TATATATATATATATATATATGGATAAATAAATATAAGGTCTATAACAGGAAAAAATGTG
TATTAACTTTTTTTTATAATTATTTTATATTTTTTCTCCATTTATTTAAATATGTTAAAA
TTATTTCCCATTTTATAAACAAAAATTAATCCATAAAATAGCAAAATAACAAAATGACAA
AATAACAAAAACAAAAAAACGCATTATATGAGTTCATTTTACACAATCCAAAGAAAAAGA
AAATATGCTTAAAATATTCATAACACACTTTTAAGCTATAAAAATATATATATTTATTTG
TGTCTATATTACCAACTCAATTTAATAGATGTGTTATGTGATTAATTCATACACAAACAT
ACAAAAATAAACACCATTTTTGAAAAGATTAATTTGAAATCATATATGTTATTATTATAT
ATTTTTATATCAATGAAATTTAAAAAAATAAAAGGGAAATCAATGTATTAAAAATAATTA
TATGTTATTTTATTTCCACAATATTTATTTATTATTTATTGTTGACCTGCAGGCATGCAA
GCTTATCGATACCGTCGACCTCGAGGGGGGGCCCGGTACCCAGCTTTTGTTCCCTTTAGT
GAGGGTTAATTGCGCGCTTGGCGTAATCATGGTCATAGCTGTTTCCTGTGTGAAATTGTT
ATCCGCTCACAATTCCACACAACATACGAGCCGGAAGCATAAAGTGTAAAGCCTGGGGTG
CCTAATGAGTGAGCTAACTCACATTAATTGCGTTGCGCTCACTGCCCGCTTTCCAGTCGG
GAAACCTGTCGTGCCAGCTGCATTAATGAATCGGCCAACGCGCGGGGAGAGGCGGTTTGC
GTATTGGGCGCTCTTCCGCTTCCTCGCTCACTGACTCGCTGCGCTCGGTCGTTCGGCTGC
GGCGAGCGGTATCAGCTCACTCAAAGGCGGTAATACGGTTATCCACAGAATCAGGGGATA
ACGCAGGAAAGAACATGTGAGCAAAAGGCCAGCAAAAGGCCAGGAACCGTAAAAAGGCCG
CGTTGCTGGCGTTTTTCCATAGGCTCCGCCCCCCTGACGAGCATCACAAAAATCGACGCT
CAAGTCAGAGGTGGCGAAACCCGACAGGACTATAAAGATACCAGGCGTTTCCCCCTGGAA
GCTCCCTCGTGCGCTCTCCTGTTCCGACCCTGCCGCTTACCGGATACCTGTCCGCCTTTC
TCCCTTCGGGAAGCGTGGCGCTTTCTCATAGCTCACGCTGTAGGTATCTCAGTTCGGTGT
AGGTCGTTCGCTCCAAGCTGGGCTGTGTGCACGAACCCCCCGTTCAGCCCGACCGCTGCG
CCTTATCCGGTAACTATCGTCTTGAGTCCAACCCGGTAAGACACGACTTATCGCCACTGG
CAGCAGCCACTGGTAACAGGATTAGCAGAGCGAGGTATGTAGGCGGTGCTACAGAGTTCT
TGAAGTGGTGGCCTAACTACGGCTACACTAGAAGGACAGTATTTGGTATCTGCGCTCTGC
TGAAGCCAGTTACCTTCGGAAAAAGAGTTGGTAGCTCTTGATCCGGCAAACAAACCACCG
CTGGTAGCGGTGGTTTTTTTGTTTGCAAGCAGCAGATTACGCGCAGAAAAAAAGGATCTC
AAGAAGATCCTTTGATCTTTTCTACGGGGTCTGACGCTCAGTGGAACGAAAACTCACGTT
AAGGGATTTTGGTCATGAGATTATCAAAAAGGATCTTCACCTAGATCCTTTTAAATTAAA
AATGAAGTTTTAAATCAATCTAAAGTATATATGAGTAAACTTGGTCTGACAGTTACCAAT
GCTTAATCAGTGAGGCACCTATCTCAGCGATCTGTCTATTTCGTTCATCCATAGTTGCCT
GACTCCCCGTCGTGTAGATAACTACGATACGGGAGGGCTTACCATCTGGCCCCAGTGCTG
CAATGATACCGCGAGACCCACGCTCACCGGCTCCAGATTTATCAGCAATAAACCAGCCAG
CCGGAAGGGCCGAGCGCAGAAGTGGTCCTGCAACTTTATCCGCCTCCATCCAGTCTATTA
ATTGTTGCCGGGAAGCTAGAGTAAGTAGTTCGCCAGTTAATAGTTTGCGCAACGTTGTTG
CCATTGCTACAGGCATCGTGGTGTCACGCTCGTCGTTTGGTATGGCTTCATTCAGCTCCG
GTTCCCAACGATCAAGGCGAGTTACATGATCCCCCATGTTGTGCAAAAAAGCGGTTAGCT
CCTTCGGTCCTCCGATCGTTGTCAGAAGTAAGTTGGCCGCAGTGTTATCACTCATGGTTA
TGGCAGCACTGCATAATTCTCTTACTGTCATGCCATCCGTAAGATGCTTTTCTGTGACTG
GTGAGTACTCAACCAAGTCATTCTGAGAATAGTGTATGCGGCGACCGAGTTGCTCTTGCC
CGGCGTCAATACGGGATAATACCGCGCCACATAGCAGAACTTTAAAAGTGCTCATCATTG
GAAAACGTTCTTCGGGGCGAAAACTCTCAAGGATCTTACCGCTGTTGAGATCCAGTTCGA
TGTAACCCACTCGTGCACCCAACTGATCTTCAGCATCTTTTACTTTCACCAGCGTTTCTG
GGTGAGCAAAAACAGGAAGGCAAAATGCCGCAAAAAAGGGAATAAGGGCGACACGGAAAT
GTTGAATACTCATACTCTTCCTTTTTCAATATTATTGAAGCATTTATCAGGGTTATTGTC
TCATGAGCGGATACATATTTGAATGTATTTAGAAAAATAAACAAATAGGGGTTCCGCGCA
CATTTCCCCGAAAAGTGCCAC
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 1AACTAGTTGTGGTGGAATACCTACTATAGC
Additional information primer 1L6455 (SpeI); PBANKA_113780 Forward
Sequence Primer 2CGGGATCCATAGTTATAACTAGCAGCATCAC
Additional information primer 2L6456 (BamHI); PBANKA_113780 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