RMgmDB - Rodent Malaria genetically modified Parasites

Summary

RMgm-743
Malaria parasiteP. berghei
Genotype
TaggedGene model (rodent): PBANKA_0701100; Gene model (P.falciparum): Not available; Gene product: conserved Plasmodium protein, unknown function
Name tag: GFP
Phenotype Asexual bloodstage;
Last modified: 2 December 2012, 15:45
  *RMgm-743
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
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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).
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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
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Name of the mutant parasite
RMgm numberRMgm-743
Principal name1397
Alternative name
Standardized name
Is the mutant parasite cloned after genetic modificationNo
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Phenotype
Asexual blood stageThe tagged-protein is exported into the cytoplasm of the host erythrocyte and shows a diffuse or patchy localization in the cytoplasm
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 GFP-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 analysed in detail.
Southern analysis of PFG-separated chromosomes (to show integration into the chromosome on which the target gene is located) and/or diagnostic PCR (for correct integration of the construct) have 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 is exported into the cytoplasm of the host erythrocyte and shows a diffuse or patchy localization in the cytoplasm.



Other mutants
RMgm-742: mutant expressing an mCherry-tagged version of  PBANKA_070110 which was introduced as a additional copy in the dSSU locus.
RMgm-502: mutant lacking expression of PBANKA_070110

  Tagged: Mutant parasite with a tagged gene
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Details of the target gene
Gene Model of Rodent Parasite PBANKA_0701100
Gene Model P. falciparum ortholog Not available
Gene productconserved Plasmodium protein, unknown function
Gene product: Alternative name
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Details of the genetic modification
Name of the tagGFP
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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GATCCATGAGTAAAGGAGAAGAACTTTTCACTGGAGTTGTCCCAATTCTTGTTGAATTAG
ATGGTGATGTTAATGGGCACAAATTTTCTGTCAGTGGAGAGGGTGAAGGTGATGCAACAT
ACGGAAAACTTACCCTTAAATTTATTTGCACTACTGGAAAACTACCTGTTCCATGGCCAA
CACTTGTCACTACTTTCGGTTATGGTGTTCAATGCTTTGCGAGATACCCAGATCATATGA
AACAGCATGACTTTTTCAAGAGTGCCATGCCCGAAGGTTATGTACAGGAAAGAACTATAT
TTTTCAAAGATGACGGGAACTACAAGACACGTGCTGAAGTCAAGTTTGAAGGTGATACCC
TTGTTAATAGAATCGAGTTAAAAGGTATTGATTTTAAAGAAGATGGAAACATTCTTGGAC
ACAAATTGGAATACAACTATAACTCACACAATGTATACATCATGGCAGACAAACAAAAGA
ATGGAATCAAAGTTAACTTCAAAATTAGACACAACATTGAAGATGGAAGCGTTCAACTAG
CAGACCATTATCAACAAAATACTCCAATTGGCGATGGCCCTGTCCTTTTACCAGACAACC
ATTACCTGTCCACACAATCTGCCCTTTCGAAAGATCCCAACGAAAAGAGAGACCACATGG
TCCTTCTTGAGTTTGTAACAGCTGCTGGGATTACACATGGCATGGATGAACTATACAAAT
AAATTCTAGAATTATTAATATATATGAATATATATACATCGTTGTATGCGTAATATACGT
GTAACTGTTTTTTTTAATTTTTATATTAACTTTTCCTTATACATTTTAATGCCATTAACT
AATAATATTTTACTTGAAAAAATTCTAAATTTTTATAACTCTAATATAAAATCTGTGAAT
AATAAAAAATTAGCATAAAGTACGAATTAAACATCGAATATTTGTTTTAAAGGGGCACAC
ATTATGTGCATATAATTTTATTCTTTGAATTTAATTTTTTGCTTATAATTAGATTAAATT
AATATAATTTTTTTATTGAAAATAAAAACTTATTTTAAAAAATTAACAAATGGAAGTTAA
TGAAAAATACGAAAAAAGTAAAGGGTTAATTCTTATATGGTCGGTACCCGAAATTGAAGG
AAAAAACATCATTTGTGTTTATATGTTTTTTTTTAATTTTTCAACTTTTTTTATTTAACT
ATGCTAATCATATTATAAACATTTTAAAAATCAAGTCCAACTATTTATGAATCATTGAAG
AGACAACATAATTATAACAGTAAAAATAATAATAATAATAATAATTTTGTCTTATTGTTT
CAAACATATTATGTTCCATAAATATCTAAATTTTTAATATGGTCATAAATTCATAGGATT
TAATTTATATAAAATATGGCTATTCATACTAGCCATTTTATGTGTGTAAAGTTTTTTTTT
TCTGAACAAAAAATAAAATGTTCATAATTTTAGCTATTTACATGCATGTGCATGCACAAA
AAAAAATATGCACACAACATACACATTTTTACAGTTATAAATACAATCAATTGGTATAAA
TATATAAGTAAGAAAAACGGGATCGAGTTAAACACAGTAGTATCTGTCACCAAAGTCACC
CAACCCTGGAACTAGATACTTGTTTTCATCTAGACCTCTGTCGAGGGCACCAGTAACAAT
TCTGACCTCTGGGAAGGCGGCATGGTATTTTTCAATCCCTTCCTTACTACAGATTAGGTT
TAAGAAGTAAATTCTCTCTGGCTTAACACCTCTCTTAATCAAGACTTCTGTAGCCATGAT
AGCACTACCACCGGTGGCCAGCATTGGGTCTAATAGGAAGACATACCTTTCAGATATATC
CTCTGGTAATTTTTCGTAGAATAACTTTGGTAAAGCAGTCTCCTCGTCCCTTTGAATTAA
AATTTTACCGATACGCACAGACCTACAACAGTCTCTTAATCCTTGCTCCATCGATTCACC
AGCTCTGACAATGGAAACACCACAGATTTTACCCATGAATGAGACACCTTCGAAGTTTTC
GTTGGTGTCAGTTTCCACAATTTGCTTTTGCACAGGTAGATGGTTCAAACCTTCTTCAAC
CAACAATCTGATGATTCTATCGGAGTAGAAAATGAAATCAGGTCTAGTTGTATTCTTATT
TCTGATGATGGTGTACAAACCCAGCAATTGGTTTGTTTGAGGTAGCAAGTAGACGTTCTT
AAATGGTTCCGAAGCCTCACCAATATCTTCAAACCAATCCTGAGGTCTTTCATCGATAAA
TTGTTTCATGATCTTTTTACACCTCTCATCGTCAACAACAACAACCTCGTGACCTCTAGT
TTGTAAATATTTCTCGCCCTTACTTTTGAAATTAACGTTCTCACCGACAACACAGCGTGG
AATACCATACATGATGATGGCACCTGTACACATGTCGCATGGAGACAGCGTCGTATACAA
AGTGGTATCTTTGTACACTTTGCCCTCTAATCTCCCACAGTTTTCCAAAGTGGAGATCTC
ACCATGTAGTGTGGCGGATCCCTTTTGAAATCTCATGTTGTGACCACGACCGAGAACACT
TCCGTCTTTGTTATTGATAAGACATCCGCCAATAGGAACACCACCCTCTTTGTAACCTAA
GGCCGCCTCCTCATAGGCAATGTCCATACCCTTCTGATCCCACTTGCTTGCCATTCCCCC
TGTCACGCTAGCACTTCCACCTCCACCAGATCCACCTCCTCCGCTAGCATCATTCTTCTC
ATATACTTCAAATTTGTACTTAATGCCTTTCTCCTCCTGGACATCAGAGAGAACACCTGG
GTATTCTGGCAGAAGTTTATATTTCTCCAAATCAATTTCTGGAAAAAACGTGTCACTTTC
AAAGTCTTGCATGATCCTTGTCACAAATAGTTTAAGATGGCCTGGGTGATTCATGGCTTC
CTTATAAACAGAACTGCCACCAACTATCCAGACCATGTCTACTTTATTTGCTAATTCTGG
TTGTTCAGTAAGTTTTAAGGCATCATCTAGACTTCTGGAAAGAAAATGAGCTCCTTGTGG
AGGTTCCTTGAGTTCTCTGCTGAGAACTAAATTAATTCTACCCTTTAAAGGTCGATTCTT
CTCAGGAATGGAGAACCAGGTCTTCTTACCCATAATCACCAGATTCTGTTTACCTTCTAC
TGAAGAGGTTGTGGTCATTCTCTGGAAATATCTAAATTCGTTCCTGAGCGGTGGCCAGGG
CAGGTCCCCGTTCTTGCCGATGCCCATGTTCTGGGACACAGCGACGATGCAGTTTAGCGA
ACCAACCATGGATCCCCCTATGTTTTATAAAATTTTTTATTTATTTATAAGCAAATATAT
ATTTTTATATATTTATACACAAAGTATATATATTTTAGGAAATATTAAAATAAATTTAAA
TTTTGAAGGCAAATGAAAAAATTACGTTTTTTGTTTCACAACAATTATATTTATTTTTTG
AATTAATTTACATATTATTTTATATTAAAATTATAAAATAAAAAAAACATTACATATATT
AATTATATTATTATATGAGGATTTTTTTATAACATAGAAATATAAATAATTACGACTAGT
TAATAAAGGGCACTTTAATTTTTATGCATAATTAGCTTATATTTAGTGCATATTTATTTT
TATACATATTTTTATATATTTTTTGATTTACATGGCGTTTTATGTTTATATGTATAACAT
TATATGCATAGTATTTTTTTGAAAATTTAATGGTATTTCAGTGGATGAAAATATTACTGG
TGCTTTGAGGGGTGAGCATTTAAAGCACAATATCTAGGATACTACAAAAAATACTTAAAG
TATGAATATTAAATTGTAAACTTAAGCATAAAGAGCTCGAAAAGAATTAAGCTGGGCTGC
AGAGGCCTGTTAACAGATCTATCGATCCGCGGGGGCCCAAGCTTGGCGTAATCATGGTCA
AGCTGTTTCCTGTGTGAAATTGTTTACCGCTCACAATTCCACACAACATACGAGCCGGAA
GCATAAAGTGTAAAGCCTGGGGTGCCTAATGAGTGAGCTAACTCACATTAATTGCGTTGC
GCTCACTGCCCGCTTTCCAGTCGGGAAACCTGTCGTGCCAGCTGCATTAATGAATCGGCC
AACGCGCGGGGAGAGGCGGTTTGCGTATTGGGCGCTCTTCCGCTTCCTCGCTCACTGACT
CGCTGCGCTCGGTCGTTCGGCTGCGGCGAGCGGTATCAGCTCACTCAAAGGCGGTAATAC
GGTTATCCACAGAATCAGGGGATAACGCAGGAAAGAACATGTGAGCAAAAGGCCAGCAAA
AGGCCAGGAACCGTAAAAAGGCCGCGTTGCTGGCGTTTTTCCATAGGCTCCGCCCCCCTG
ACGAGCATCACAAAAATCGACGCTCAAGTCAGAGGTGGCGAAACCCGACAGGACTATAAA
GATACCAGGCGTTTCCCCCTGGAAGCTCCCTCGTGCGCTCTCCTGTTCCGACCCTGCCGC
TTACCGGATACCTGTCCGCCTTTCTCCCTTCGGGAAGCGTGGCGCTTTCTCAATGCTCAC
GCTGTAGGTATCTCAGTTCGGTGTAGGTCGTTCGCTCCAAGCTGGGCTGTGTGCACGAAC
CCCCCGTTCAGCCCGACCGCTGCGCCTTATCCGGTAACTATCGTCTTGAGTCCAACCCGG
TAAGACACGACTTATCGCCACTGGCAGCAGCCACTGGTAACAGGATTAGCAGAGCGAGGT
ATGTAGGCGGTGCTACAGAGTTCTTGAAGTGGTGGCCTAACTACGGCTACACTAGAAGGA
CAGTATTTGGTATCTGCGCTCTGCTGAAGCCAGTTACCTTCGGAAAAAGAGTTGGTAGCT
CTTGATCCGGCAAACAAACCACCGCTGGTAGCGGTGGTTTTTTTGTTTGCAAGCAGCAGA
TTACGCGCAGAAAAAAAGGATCTCAAGAAGATCCTTTGATCTTTTCTACGGGGTCTGACG
CTCAGTGGAACGAAAACTCACGTTAAGGGATTTTGGTCATGAGATTATCAAAAAGGATCT
TCACCTAGATCCTTTTAAATTAAAAATGAAGTTTTAAATCAATCTAAAGTATATATGAGT
AAACTTGGTCTGACAGTTACCAATGCTTAATCAGTGAGGCACCTATCTCAGCGATCTGTC
TATTTCGTTCATCCATAGTTGCCTGACTCCCCGTCGTGTAGATAACTACGATACGGGAGG
GCTTACCATCTGGCCCCAGTGCTGCAATGATACCGCGAGACCCACGCTCACCGGCTCCAG
ATTTATCAGCAATAAACCAGCCAGCCGGAAGGGCCGAGCGCAGAAGTGGTCCTGCAACTT
TATCCGCCTCCATCCAGTCTATTAATTGTTGCCGGGAAGCTAGAGTAAGTAGTTCGCCAG
TTAATAGTTTGCGCAACGTTGTTGCCATTGCTACAGGCATCGTGGTGTCACGCTCGTCGT
TTGGTATGGCTTCATTCAGCTCCGGTTCCCAACGATCAAGGCGAGTTACATGATCCCCCA
TGTTGTGCAAAAAAGCGGTTAGCTCCTTCGGTCCTCCGATCGTTGTCAGAAGTAAGTTGG
CCGCAGTGTTATCACTCATGGTTATGGCAGCACTGCATAATTCTCTTACTGTCATGCCAT
CCGTAAGATGCTTTTCTGTGACTGGTGAGTACTCAACCAAGTCATTCTGAGAATAGTGTA
TGCGGCGACCGAGTTGCTCTTGCCCGGCGTCAATACGGGATAATACCGCGCCACATAGCA
GAACTTTAAAAGTGCTCATCATTGGAAAACGTTCTTCGGGGCGAAAACTCTCAAGGATCT
TACCGCTGTTGAGATCCAGTTCGATGTAACCCACTCGTGCACCCAACTGATCTTCAGCAT
CTTTTACTTTCACCAGCGTTTCTGGGTGAGCAAAAACAGGAAGGCAAAATGCCGCAAAAA
AGGGAATAAGGGCGACACGGAAATGTTGAATACTCATACTCTTCCTTTTTCAATATTATT
GAAGCATTTATCAGGGTTATTGTCTCATGAGCGGATACATATTTGAATGTATTTAGAAAA
ATAAACAAATAGGGGTTCCGCGCACATTTCCCCGAAAAGTGCCACCTGACGTCTAAGAAA
CCATTATTATCATGACATTAACCTATAAAAATAGGCGTATCACGAGGCCCTTTCGTCTCG
CGCGTTTCGGTGATGACGGTGAAAACCTCTGACACATGCAGCTCCCGGAGACGGTCACAG
CTTGTCTGTAAGCGGATGCCGGGAGCAGACAAGCCCGTCAGGGCGCGTCAGCGGGTGTTG
GCGGGTGTCGGGGCTGGCTTAACTATGCGGCATCAGAGCAGATTGTACTGAGAGTGCACC
ATATGCGGTGTGAAATACCGCACAGATGCGTAAGGAGAAAATACCGCATCAGGCGCCATT
CGCCATTCAGGCTGCGCAACTGTTGGGAAGGGCGATCGGTGCGGGCCTCTTCGCTATTAC
GCCAGCTGGCGAAAGGGGGATGTGCTGCAAGGCGATTAAGTTGGGTAACGCCAGGGTTTT
CCCAGTCACGACGTTGTAAAACGACGGCCAGTGAATTCGCGGCCGCTTAATATAATATGC
AATAGTAACACAATTATTATATATGTTTTATATATATGAAAAAAATGCATACGTCATCTA
AATATTTATTTTTCTTCATCAATAAAATATATCCTAAAATATTAGGGTACAAAAGAAAAT
TTTTATTTTAATATATTATTATTTTATCCCACTTTCGTATTTACTTATTATTAATATATA
ATAATTACAAATAATGGTATTTTTATCATCAAGATTTATGTTTATTTTTATTATGCACAA
AAAATAATATATAATAAAATTGTATAAGATACATATTTACATGAGTAGTATTAGTTTTAC
TATTTTACTATAGTTTTTATTCATTTATAATTTTTTTATATTGTAAAATATTATTATTAC
TATTATTATTTTATTAGAGTTTTGTAAATATTTATTTAAATTCCTAATATTTTTATTTAC
ATTATATATAGCTAAAAATAATATTAATGAATTTTTAAGCTTCAAAAGGCATTTAATATT
TATCCTATTATTTTGTTATATTAATATTTAACTTTTTTAATTATTATATTCTACAATATG
AAGCACAAATTGCATACCTTTTTTGTTATCATATGCTTAGTCGTAGTATTGTATTGTGGT
GGTATAGAGGGAAAACAAAATAATAATATAAAAAAGGATATGTGGATATCCCGATTTTTT
ACTCATAAAAACGGCAAATATGCGAGAAATGAAAAAACAGTAGAAAACGCGGAAGGTGGT
AATAAAAACAATGAAATATTAGATGATCTATATAATGAAGATATAGGTATGGTATTTTAA
TATTAAAGATATGCGCATATATTATACATGAATTCCTCATAATTTGGTAGAAAAAAAGTT
TACAAACAATATAAATGCTTATCCAACTTATTATTTTTTACACAATGTTGTTTTATTTGA
TAATACTATGCATTATACACATTTTTTATTTATATTTTTTTAAACCTTCAGATGAGGTAG
TCAAAAATATCCTTAAAAGTGAAAATGTTGATATTCAAACTTTGAAGATTATAAAAGAAC
TTGATAAAAAAAAAAATGAAGTAATTATTGCCAATAAACAAAAAAATTTATATAAATCTG
GCGCATTTGGTATGAGTGTAATATTTCTTAGCTTTGCTTTGCGTGAACTCGCAGAACTTT
TAGTAAACTATTATCTAGACAAACATCATAATACTAAAAATGCACTTAACCGTTTGATTA
CGAACAAATGGAAAG
Restriction sites to linearize plasmid EcoRV
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 1AGGATCCTTTCCATTTGTTCGTAATCAAAC
Additional information primer 14371
Sequence Primer 2aagatctGCGGCCGCTTAATATAATATGCAATAGTAACAC
Additional information primer 24372
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
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Conceptual design of the database: Dr. C.J. Janse (Leiden Malaria Research Group, Leiden, The Netherlands).
Technical design and realization: S. Mededovic and A. van Wigcheren