Summary

RMgm-714
Malaria parasiteP. berghei
Genotype
TaggedGene model (rodent): PBANKA_1238800; Gene model (P.falciparum): PF3D7_0524000; Gene product: karyopherin beta (KASbeta)
Name tag: mCherry
Phenotype Asexual bloodstage;
Last modified: 2 December 2012, 15:42
  *RMgm-714
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-714
Principal name1956
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


  Tagged: Mutant parasite with a tagged gene
Details of the target gene
Gene Model of Rodent Parasite PBANKA_1238800
Gene Model P. falciparum ortholog PF3D7_0524000
Gene productkaryopherin beta
Gene product: Alternative nameKASbeta
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
CCGCGGTGGCGGCCGCTCTAGAACTAGTCAGCAAGAATAATATCAGATCAAACAGATAAT
AATAAATCTATGCTATTAGCTATACTTACAACATCTGCTGAAAAAGTATTAAAAAGTTTA
TCAGATACAAAATTAGATGATAATTATGAATATGTATTAGATATAATGATTATCGAATCT
AATGGTTTATATATGTGCTTACAAAAAGCCGGTGCTAATATATTACCTAACAATACATTA
AAAATGTTTTTTAATGAAATATTTAAATTATTACAATATTCAACAGATAGAAGAGTCATT
TATAATCAAAAAAAAAATAATGAAGATGTCGATGATGATGAATTATTAATTATAGACAGA
GAAGAAGAGCTAGAACAAACATATCGAACAAACTTATTAGACATATTGGGAGTATTAATA
AAACATCACACAAATCAATTCTTAAGTACATGTTGTGATATATGTATTACATTTATAAAT
AATTATTTAAACTCACCACATGCAGAAGATATTGCATTGGCTTTATATGTTTGTGATGAT
TTGTTAGAATTTTTGCAAGATAATAGTGTATGCTTGTGGGAATATTTTATGAACCCATTA
TTATTAAATATTAATCATACAGATCATAAAGTTAAACAAGCAGCTTGTTATGGTGTTATA
CAAGCAAATAAAATAGAAGCATTTGGAAAATATGCAAATTTGGCCATAGAATATTTATTA
AAACTATTACATCAAACTCCCCCAAGTAAAAAACCCAAGGAATACATATCTGCTATTGAT
AATGCTGTAGCTGCATTAGGAGATGTTGTCTTAATGCACACATCCAAATTCAATAATTCA
GAAGAATTAATTAAGTTGTGGTTAAATAATTTACCAATTAAAGAAGATGAATCAGAAGGA
AGACGAGTGCACAAAAACTTAATCGATTTAGTTTCTCAAAATCACCCACTTTTATTCGGA
AAAGATAATTCTAATATTGGAAAAATTATAGAAATATTCCTATCAATATATGAAACAGAA
TTTTCTGATTCAGATTGTAACAAAAAAATAGTATCCTTGATAAGTTCTTTAGATCAATCT
TATTTAAGCAATTTGGCTTCCACATCTCTCACCAACAAACAATCCAAAAAATTGAATCAC
ATTATGAACGCCTCTAGAAAGGGATCCGGTGGAGGTGGAAGATCCATGGTGAGCAAGGGC
GAGGAGGATAACATGGCCATCATCAAGGAGTTCATGCGCTTCAAGGTGCACATGGAGGGC
TCCGTGAACGGCCACGAGTTCGAGATCGAGGGCGAGGGCGAGGGCCGCCCCTACGAGGGC
ACCCAGACCGCCAAGCTGAAGGTGACCAAGGGTGGCCCCCTGCCCTTCGCCTGGGACATC
CTGTCCCCTCAGTTCATGTACGGCTCCAAGGCCTACGTGAAGCACCCCGCCGACATCCCC
GACTACTTGAAGCTGTCCTTCCCCGAGGGCTTCAAGTGGGAGCGCGTGATGAACTTCGAG
GACGGCGGCGTGGTGACCGTGACCCAGGACTCCTCCCTGCAGGACGGCGAGTTCATCTAC
AAGGTGAAGCTGCGCGGCACCAACTTCCCCTCCGACGGCCCCGTAATGCAGAAGAAGACC
ATGGGCTGGGAGGCCTCCTCCGAGCGGATGTACCCCGAGGACGGCGCCCTGAAGGGCGAG
ATCAAGCAGAGGCTGAAGCTGAAGGACGGCGGCCACTACGACGCTGAGGTCAAGACCACC
TACAAGGCCAAGAAGCCCGTGCAGCTGCCCGGCGCCTACAACGTCAACATCAAGTTGGAC
ATCACCTCCCACAACGAGGACTACACCATCGTGGAACAGTACGAACGCGCCGAGGGCCGC
CACTCCACCGGCGGCATGGACGAGCTGTACAAGTAATCTAGAATTATTAATATATATGAA
TATATATACATCGTTGTATGCGTAATATACGTGTAACTGTTTTTTTTAATTTTTATATTA
ACTTTTCCTTATACATTTTAATGCCATTAACTAATAATATTTTACTTGAAAAAATTCTAA
ATTTTTATAACTCTAATATAAAATCTGTGAATAATAAAAAATTAGCATAAAGTACGAATT
AAACATCGAATATTTGTTTTAAAGGGGCACACATTATGTGCATATAATTTTATTCTTTGA
ATTTAATTTTTTGCTTATAATTAGATTAAATTAATATAATTTTTTTATTGAAAATAAAAA
CTTATTTTAAAAAATTAACAAATGGAAGTTAATGAAAAATACGAAAAAAGTAAAGGGTTA
ATTCTTATATGGTCGGTACCGAGCTCGAATTCGATATCGAAATTGAAGGAAAAAACATCA
TTTGTGTTTATATGTTTTTTTTTAATTTTTCAACTTTTTTTATTTAACTATGCTAATCAT
ATTATAAACATTTTAAAAATCAAGTCCAACTATTTATGAATCATTGAAGAGACAACATAA
TTATAACAGTAAAAATAATAATAATAATAATAATTTTGTCTTATTGTTTCAAACATATTA
TGTTCCATAAATATCTAAATTTTTAATATGGTCATAAATTCATAGGATTTAATTTATATA
AAATATGGCTATTCATACTAGCCATTTTATGTGTGTAAAGTTTTTTTTTTCTGAACAAAA
AATAAAATGTTCATAATTTTAGCTATTTACATGCATGTGCATGCACAAAAAAAAATATGC
ACACAACATACACATTTTTACAGTTATAAATACAATCAATTGGTATAAATATATAAGTAA
GAAAAACGGGATCAAAGCTAGCTTCTGTATTTCCGCTAGACAGCCATCTCCATCTGGATT
CGTCCGTGCGGGACGTAGCCCACGACCTCAAAATCCTCGGCGGTGAAATCGTCGATTTCC
TTGATGCGTTCCTTGTTGAGGATGTTCACAATGGGGAACGGTCTCGGTTCTCTCCGCAGC
TGCTCTTTTAAAGCCTCGACATGGTTCGTGTAGACATGCGTGTTCCCCATGAAGTGAATG
AACTCCTTAGGTTTTAGGTTGCAGACGTGTGCAACCATGAGCGTCAAAAGCGAATAGGAA
GCGATGTTGAAGGGGACGCCGAGGCCGACATCGCACGACCGCTGATACATGATGCACGAC
AGCTCCTTCTGGTCGTTCACGTAGAACTGGCACAACAAGTGACAAGGCGGCAGCGCCATT
TCGTCCAGCGCTGCAGGATTCCAGGCAGTCATGAGCATGCGACGATCTGTTGGATTCGTT
CTCAGCATCTGGATCACATTCTTCAGCTGGTCGACGCCCTGCCCTGTGTAGTCTGTGTGC
ATGTCTTTGTATGCCGCGCCGAAGTGTCTCCACTGGAAGCCGTAGCCCGGGCCGATGTCT
CCGACCTCTCGGTGGGGGAGATTGCGCGAATCGAGGAACTCGCGTGTCACATTCTTGTCC
CAGATCTTCACGCCCTTCTCAGAAAGATGGTTTGCGTTCGTGTCGCCGCGAATGAACCAC
AGCAACTCTTCGAGGACCCCTTTCCAGAACACACGCTTTGTGGTGAGAAGTGGAAAGGCC
TGATCCAGCGAGTAGCGCATAGTGCAGCCGAATTTGGAGATGACACCAACGCCCGTTCGG
TCATCCATTGTCCTTCCATTGTTAATAATGTCGGCAATGAGATCAAGGTACTGGAACTCT
TCATGGCCTCTAAAGTGAACATGCGGAACGGCCCGAATCAGTTCCTTTTGCTCGCGTTTT
TTCCGGTCTTCTTCGTCCATCCACGCCAACACCGGGGCAATGGCTGCGGCCGAAGAAGGA
GCCTGCAACCCGTGCACGGGAGTTGTCTCCCTCGTGGACGTCAAGGAGCTCATTGCGTTG
CTCGGTTCCGCAGTGGCTGCGTCGTCAGTCTTCCTTCTCTTCTCGAGAACCACAAAGTCG
TAGGGAACCCCGTTGTCTGAGAAGGTCTTGGAAATGAAGATGGGTCGATACGTCGCTTCA
TTGTCCTTCTCTCTTCCGAGCTCCGGACAAAAGGGAACGAACACAGACTCGGCAGGAGCT
GCAGCCTGCGCAGCAGTTGATTTGTTTGAAAGAATGTCATCTCCGGGGAACGCAGGGAAG
AAAACGTCGCACGGAAACTCGCGGGCTACACGCGTGATGTACAGGTGAGAGGCAACGCCC
AGAGACAGCGCTGCCTCGTACAGTCCCGCTCCTCCCACGACAAAAATCTGGTCGACAGAA
TCCTTGTACTCTTCCTCCAGAAGGCTGAGAGCTGCTGGGAGTGAAGCACAGACTCGGACG
CGCTGCTGGCCTTCAGCTTGAGGCTTCTCCGCCGCAATGTCTTCTTCTTTGAGGGAAGAG
GAAACGACGATGTTCAATCTGTCCACGAGGGGTCTAAACTTTCGAGGCATGCTTTCCCAG
GTTTTCCGTCCCATGACAACGGCGTTGAATCTCTTGCCGACTGATGGAGAGGGAAGTCCA
GAGTCGCCCGTCTTTGCAAATTTCCTGGGAAGCCACCCGTTCAGGCGACTGGCTTCTTCG
GGCGTCGTTTTTGTCACACGAGAAAAGTGTTTGAAATCTGTGGTCAAGTGGGGCCACGGG
AGGCCGTTGTTGATGCCGATGCCCCTCTTGGGGGTCATCGCGACGACCAGACACACCGGT
TTATGCATTGATCCTTTGTAACATTTAGGTGTGTATTTATATATATAAGCAATATACAAA
TGAAAATACTATTATAAAACGGAAACTCAAAAAATGTTGAAACAAATAGTCAATTTTTAA
AATGAATGTGTAAATTGTTTAAGTTTTATGACAAAAAAAGGAAAAGTGTATATAAAATAA
AAATTGGAAAAAATTTATATATTACATGCCATATAATACCATTTATTTACTATTTATATC
TTAATATAAAAAATTGTAGCAAAAATATATAATCGTATTTTATATATTTAATATTATATA
ACCCCTTAAAAAAAAAAATTTAATTGAAAAAATTTTAATTTATGCACTTTTTTATTGTAT
TATATATATTCCGTTATTTTAATTACACATAAACTTAATAGAAAAATTAATTCGCTGCTA
AATAAAAATTATAGGGCTATATATGATGGGATCATATTCTATCTTTCTCTTTTTCAAATY
CTTTATATTTTAGAATACAAATCGCTTTAAACAATAAAAAAGAGGTATATCCTACCCGAA
TTGACACACACAATAAATAATAAATATATAAATAATAAAAATATAAATATATAAATTATA
AATATATAAATTATATATATAAATATATTAACATGTGTGTAATTTGTTAAAATTGATATT
AAACAGCTTTGCATGGGGAAAGAGTGGCTTTTATAAGCGAATCATATAATACAGTGTTAA
TTTTAATATATAAAATATAATTTCCTTAAATTGTTATTTAACATTTTTAATTGCCTACCC
ARGGTAATATAATATCCAAATGACAATCAAAGATAAATGAAATACCGCTCCATTTTTCCT
ATATATCCATTTTCATTTTGTGTAAATGTTCATAAAATTTTATCATATACTCATAAAAAA
CAGGGGAAAAATGAAAAAATGAACGTCCACCCCGTGTGAATATGCTCATTTTGTATACTA
TACATTTTTTTTTATATAATTTATATTTTCCAAATAAAAAAAATGTTTGTACATGTATAT
ATCAATATTGTGTTATGGCATGATGAAAAACAATTATTTTATTCGTTTTTAAATTTGTAG
AATTTCCCTAAATTATTAATGATTTCATATTATAATATATGTATCTATATATTTTTAATG
TGTAATTCAAAATGGCGAAAAAGAAACAAACCATACTTTATAAGCAACTCTCTTTGAGCA
CTTACCATAAAAAATTATACTACTAAATTCAAATTAAATAAAAAAATATATATTATAAAA
TTGGCTTTAATTATTTATAAGCACTTGCTTTGTTTTACTCAAAAAGTATTGGCATAAGTA
TAAACATAAAACCAATTTTTCACAAAATATATTATATGTCCATACAACTATATCCGAACY
TTCAAAACACATCCAAATTTCCATTTACCACATGCAAGCAAAAGGATATTAAATAAAAAA
AAATTATTTATATTTTCATGATCACTTAAATAATATTTAAAAAAAAAAGAGACCATATAT
AAAATGTATAATTTAAATATAGCTAACTTATTAAAAAAAATGAATATTGAGCATATATCC
ATAATTTAAAACACTTCTATTTTATGTTATTTGATATATAAGTCGATAAATTATTGCATG
TATGCATATAGACAGATTAATATACACCAAAATAATAAAAATTATATATAGTTCATATAT
TATTCTGAATAATAAATTAGCTTTCCTCTTAATTCACTTGGTATGTATATATATATATAT
ATATATATATATATATATATGGATAAATAAATATAAGGTCTATAACAGGAAAAAATGTGT
ATTAACTTTTTTTTATAATTATTTTATATTTTTTCTCCATTTATTTAAATATGTTAAAAT
TATTTCCCATTTTATAAACAAAAATTAATCCATAAAATAGCAAAATAACAAAATGACAAA
ATAACAAAAACAAAAAAACGCATTATATGAGTTCATTTTACACAATCCAAAGAAAAAGAA
AATATGCTTAAAATATTCATAACACACTTTTAAGCTATAAAAATATATATATTTATTTGT
GTCTATATTACCAACTCAATTTAATAGATGTGTTATGTGATTAATTCATACACAAACATA
CAAAAATAAACACCATTTTTGAAAAGATTAATTTGAAATCATATATGTTATTATTATATA
TTTTTATATCAATGAAATTTAAAAAAATAAAAGGGAAATCAATGTATTAAAAATAATTAT
ATGTTATTTTATTTCCACAATATTTATTTATTATTTATTGTTGACCTGCAGGCATGCAAG
CTTATCGATACCGTCGACCTCGAGGGGGGGCCCGGTACCCAGCTTTTGTTCCCTTTAGTG
AGGGTTAATTGCGCGCTTGGCGTAATCATGGTCATAGCTGTTTCCTGTGTGAAATTGTTA
TCCGCTCACAATTCCACACAACATACGAGCCGGAAGCATAAAGTGTAAAGCCTGGGGTGC
CTAATGAGTGAGCTAACTCACATTAATTGCGTTGCGCTCACTGCCCGCTTTCCAGTCGGG
AAACCTGTCGTGCCAGCTGCATTAATGAATCGGCCAACGCGCGGGGAGAGGCGGTTTGCG
TATTGGGCGCTCTTCCGCTTCCTCGCTCACTGACTCGCTGCGCTCGGTCGTTCGGCTGCG
GCGAGCGGTATCAGCTCACTCAAAGGCGGTAATACGGTTATCCACAGAATCAGGGGATAA
CGCAGGAAAGAACATGTGAGCAAAAGGCCAGCAAAAGGCCAGGAACCGTAAAAAGGCCGC
GTTGCTGGCGTTTTTCCATAGGCTCCGCCCCCCTGACGAGCATCACAAAAATCGACGCTC
AAGTCAGAGGTGGCGAAACCCGACAGGACTATAAAGATACCAGGCGTTTCCCCCTGGAAG
CTCCCTCGTGCGCTCTCCTGTTCCGACCCTGCCGCTTACCGGATACCTGTCCGCCTTTCT
CCCTTCGGGAAGCGTGGCGCTTTCTCATAGCTCACGCTGTAGGTATCTCAGTTCGGTGTA
GGTCGTTCGCTCCAAGCTGGGCTGTGTGCACGAACCCCCCGTTCAGCCCGACCGCTGCGC
CTTATCCGGTAACTATCGTCTTGAGTCCAACCCGGTAAGACACGACTTATCGCCACTGGC
AGCAGCCACTGGTAACAGGATTAGCAGAGCGAGGTATGTAGGCGGTGCTACAGAGTTCTT
GAAGTGGTGGCCTAACTACGGCTACACTAGAAGGACAGTATTTGGTATCTGCGCTCTGCT
GAAGCCAGTTACCTTCGGAAAAAGAGTTGGTAGCTCTTGATCCGGCAAACAAACCACCGC
TGGTAGCGGTGGTTTTTTTGTTTGCAAGCAGCAGATTACGCGCAGAAAAAAAGGATCTCA
AGAAGATCCTTTGATCTTTTCTACGGGGTCTGACGCTCAGTGGAACGAAAACTCACGTTA
AGGGATTTTGGTCATGAGATTATCAAAAAGGATCTTCACCTAGATCCTTTTAAATTAAAA
ATGAAGTTTTAAATCAATCTAAAGTATATATGAGTAAACTTGGTCTGACAGTTACCAATG
CTTAATCAGTGAGGCACCTATCTCAGCGATCTGTCTATTTCGTTCATCCATAGTTGCCTG
ACTCCCCGTCGTGTAGATAACTACGATACGGGAGGGCTTACCATCTGGCCCCAGTGCTGC
AATGATACCGCGAGACCCACGCTCACCGGCTCCAGATTTATCAGCAATAAACCAGCCAGC
CGGAAGGGCCGAGCGCAGAAGTGGTCCTGCAACTTTATCCGCCTCCATCCAGTCTATTAA
TTGTTGCCGGGAAGCTAGAGTAAGTAGTTCGCCAGTTAATAGTTTGCGCAACGTTGTTGC
CATTGCTACAGGCATCGTGGTGTCACGCTCGTCGTTTGGTATGGCTTCATTCAGCTCCGG
TTCCCAACGATCAAGGCGAGTTACATGATCCCCCATGTTGTGCAAAAAAGCGGTTAGCTC
CTTCGGTCCTCCGATCGTTGTCAGAAGTAAGTTGGCCGCAGTGTTATCACTCATGGTTAT
GGCAGCACTGCATAATTCTCTTACTGTCATGCCATCCGTAAGATGCTTTTCTGTGACTGG
TGAGTACTCAACCAAGTCATTCTGAGAATAGTGTATGCGGCGACCGAGTTGCTCTTGCCC
GGCGTCAATACGGGATAATACCGCGCCACATAGCAGAACTTTAAAAGTGCTCATCATTGG
AAAACGTTCTTCGGGGCGAAAACTCTCAAGGATCTTACCGCTGTTGAGATCCAGTTCGAT
GTAACCCACTCGTGCACCCAACTGATCTTCAGCATCTTTTACTTTCACCAGCGTTTCTGG
GTGAGCAAAAACAGGAAGGCAAAATGCCGCAAAAAAGGGAATAAGGGCGACACGGAAATG
TTGAATACTCATACTCTTCCTTTTTCAATATTATTGAAGCATTTATCAGGGTTATTGTCT
CATGAGCGGATACATATTTGAATGTATTTAGAAAAATAAACAAATAGGGGTTCCGCGCAC
ATTTCCCCGAAAAGTGCCAC
Restriction sites to linearize plasmid PacI
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 1GGACTAGTCAGCAAGAATAATATCAGATCAAACAG
Additional information primer 1L6657 (SpeI); PBANKA_123880 Forward
Sequence Primer 2CGGGATCCCTTTCTAGAGGCGTTCATAATGTG
Additional information primer 2L6658 (BamHI); PBANKA_123880 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