RMgmDB - Rodent Malaria genetically modified Parasites

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Summary

RMgm-499
Malaria parasiteP. berghei
Genotype
DisruptedGene model (rodent): PBANKA_0700700; Gene model (P.falciparum): PF3D7_0301800; Gene product: Plasmodium exported protein, unknown function
PhenotypeNo phenotype has been described
Last modified: 6 November 2015, 15:00
  *RMgm-499
Successful modificationThe parasite was generated by the genetic modification
The mutant contains the following genetic modification(s) Gene disruption
Reference (PubMed-PMID number) Reference 1 (PMID number) : 22184632
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 1037m1f1mocl1 (RMgm-32)
Other information parent lineP. berghei ANKA 1037m1f1mocl1 (1037cl1; RMgm-32) is a reference ANKA mutant line which expresses GFP-luciferase under control of a schizont-specific promoter (PubMed: PMID: 20019192). It does not contain a drug-selectable marker and has been selected by FACS sorting based on GFP expression
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The mutant parasite was generated by
Name PI/ResearcherJ. Fonager, E.M. Pasini, C.J. Janse, B.M Franke-Fayard
Name Group/DepartmentLeiden Malaria Research Group
Name InstituteLeiden University Medical Center (LUMC)
CityLeiden
CountryThe Netherlands
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Name of the mutant parasite
RMgm numberRMgm-499
Principal nameexp. 1485
Alternative name
Standardized name
Is the mutant parasite cloned after genetic modificationNo
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Phenotype
Asexual blood stageNot different from wild type
Gametocyte/GameteNot tested
Fertilization and ookineteNot tested
OocystNot tested
SporozoiteNot tested
Liver stageNot tested
Additional remarks phenotype

A pyrimethamine-resistant parasite population was selected after transfection with a construct for targeted gene disruption by double cross-over homologous recombination. The primers of the two target sequences of the gene are provided below.

The genotype of the parasites has not been analysed in detail.
Diagnostic PCR (for correct integration of the construct) and/or Southern analysis of PFG-separated chromosomes (to show integration into the chromosome on which the target gene is located) have been performed (see below). This analysis provided evidence for correct disruption of the gene indicating that it has no essential function during asexual blood stage development. However, no further analyses on the parasite population/cloned lines have been performed to show absence of transcripts or protein .

The phenotype of the selected parasite population/cloned lines has not been analysed in detail.
No obvious growth defects of the asexual blood stages of the selected parasite populations/cloned lines have been observed.

The gene has been selected for targeted gene disruption based on its presence in a proteome of membranes of infected red blood cells

  Disrupted: Mutant parasite with a disrupted gene
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Details of the target gene
Gene Model of Rodent Parasite PBANKA_0700700
Gene Model P. falciparum ortholog PF3D7_0301800
Gene productPlasmodium exported protein, unknown function
Gene product: Alternative name
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Details of the genetic modification
Inducable system usedNo
Additional remarks inducable system
Type of plasmid/construct used(Linear) plasmid double 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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ATCCTTACTAAAGTGGTTCGCGACGAATGCCAAAATTGGGTTACGACTCAATTAATGGCT
TAATCAACGATTATATTAATTTGTTATTTTTAATGATACGAACGTAATGCCTTTATTATA
AAAAAATTCCTAAAGCAAAAAATATAACGTGGTTCTATAATTATTAAATAATAATATAAA
GCTAGAGAATAAATAAGTAAAATAAGTTATAATTATTATACAATTCTGCCATATGCTTAT
GAGCGTATAATGCATGTTAATATAGTATAAAATAATGAACTAATGGAATAACGTCGACAA
TCTGTATATATATGCATATGCATATTTTGATTAGATTAATTTTATATTTGTTTTAATTCG
CTATATGCCTTGATTTATTATTTTTTGTATTACTCTAGTTATATCATTTTTCAGTATCAT
TATTATTCTTAAATGTATTTCATAGCGAATTATTTTTTTTACATATGAATAAGATATAAT
TTATATATAGAATCATTACTTAAATATTCCTTTTTTTATACTTTTGCTTTTTTGTGTTTA
CGCATTAAAACTTATTATGGAATTCCTGCAGCCCGGGGGATCCACTAGTTCTAGAGCGGC
CGCCACCGCGGTGGAGCTCCAATTCGCCCTATAGTGAGTCGTATTACAATTCACTGGCCG
TCGTTTTACAACGTCGTGACTGGGAAAACCCTGGCGTTACCCAACTTAATCGCCTTGCAG
CACATCCCCCTTTCGCCAGCTGGCGTAATAGCGAAGAGGCCCGCACCGATCGCCCTTCCC
AACAGTTGCGCAGCCTGAATGGCGAATGGGACGCGCCCTGTAGCGGCGCATTAAGCGCGG
CGGGTGTGGTGGTTACGCGCAGCGTGACCGCTACACTTGCCAGCGCCCTAGCGCCCGCTC
CTTTCGCTTTCTTCCCTTCCTTTCTCGCCACGTTCGCCGGCTTTCCCCGTCAAGCTCTAA
ATCGGGGGCTCCCTTTAGGGTTCCGATTTAGTGCTTTACGGCACCTCGACCCCAAAAAAC
TTGATTAGGGTGATGGTTCACGTAGTGGGCCATCGCCCTGATAGACGGTTTTTCGCCCTT
TGACGTTGGAGTCCACGTTCTTTAATAGTGGACTCTTGTTCCAAACTGGAACAACACTCA
ACCCTATCTCGGTCTATTCTTTTGATTTATAAGGGATTTTGCCGATTTCGGCCTATTGGT
TAAAAAATGAGCTGATTTAACAAAAATTTAACGCGAATTTTAACAAAATATTAACGCTTA
CAATTTAGGTGGCACTTTTCGGGGAAATGTGCGCGGAACCCCTATTTGTTTATTTTTCTA
AATACATTCAAATATGTATCCGCTCATGAGACAATAACCCTGATAAATGCTTCAATAATA
TTGAAAAAGGAAGAGTATGAGTATTCAACATTTCCGTGTCGCCCTTATTCCCTTTTTTGC
GGCATTTTGCCTTCCTGTTTTTGCTCACCCAGAAACGCTGGTGAAAGTAAAAGATGCTGA
AGATCAGTTGGGTGCACGAGTGGGTTACATCGAACTGGATCTCAACAGCGGTAAGATCCT
TGAGAGTTTTCGCCCCGAAGAACGTTTTCCAATGATGAGCACTTTTAAAGTTCTGCTATG
TGGCGCGGTATTATCCCGTATTGACGCCGGGCAAGAGCAACTCGGTCGCCGCATACACTA
TTCTCAGAATGACTTGGTTGAGTACTCACCAGTCACAGAAAAGCATCTTACGGATGGCAT
GACAGTAAGAGAATTATGCAGTGCTGCCATAACCATGAGTGATAACACTGCGGCCAACTT
ACTTCTGACAACGATCGGAGGACCGAAGGAGCTAACCGCTTTTTTGCACAACATGGGGGA
TCATGTAACTCGCCTTGATCGTTGGGAACCGGAGCTGAATGAAGCCATACCAAACGACGA
GCGTGACACCACGATGCCTGTAGCAATGGCAACAACGTTGCGCAAACTATTAACTGGCGA
ACTACTTACTCTAGCTTCCCGGCAACAATTAATAGACTGGATGGAGGCGGATAAAGTTGC
AGGACCACTTCTGCGCTCGGCCCTTCCGGCTGGCTGGTTTATTGCTGATAAATCTGGAGC
CGGTGAGCGTGGGTCTCGCGGTATCATTGCAGCACTGGGGCCAGATGGTAAGCCCTCCCG
TATCGTAGTTATCTACACGACGGGGAGTCAGGCAACTATGGATGAACGAAATAGACAGAT
CGCTGAGATAGGTGCCTCACTGATTAAGCATTGGTAACTGTCAGACCAAGTTTACTCATA
TATACTTTAGATTGATTTAAAACTTCATTTTTAATTTAAAAGGATCTAGGTGAAGATCCT
TTTTGATAATCTCATGACCAAAATCCCTTAACGTGAGTTTTCGTTCCACTGAGCGTCAGA
CCCCGTAGAAAAGATCAAAGGATCTTCTTGAGATCCTTTTTTTCTGCGCGTAATCTGCTG
CTTGCAAACAAAAAAACCACCGCTACCAGCGGTGGTTTGTTTGCCGGATCAAGAGCTACC
AACTCTTTTTCCGAAGGTAACTGGCTTCAGCAGAGCGCAGATACCAAATACTGTCCTTCT
AGTGTAGCCGTAGTTAGGCCACCACTTCAAGAACTCTGTAGCACCGCCTACATACCTCGC
TCTGCTAATCCTGTTACCAGTGGCTGCTGCCAGTGGCGATAAGTCGTGTCTTACCGGGTT
GGACTCAAGACGATAGTTACCGGATAAGGCGCAGCGGTCGGGCTGAACGGGGGGTTCGTG
CACACAGCCCAGCTTGGAGCGAACGACCTACACCGAACTGAGATACCTACAGCGTGAGCT
ATGAGAAAGCGCCACGCTTCCCGAAGGGAGAAAGGCGGACAGGTATCCGGTAAGCGGCAG
GGTCGGAACAGGAGAGCGCACGAGGGAGCTTCCAGGGGGAAACGCCTGGTATCTTTATAG
TCCTGTCGGGTTTCGCCACCTCTGACTTGAGCGTCGATTTTTGTGATGCTCGTCAGGGGG
GCGGAGCCTATGGAAAAACGCCAGCAACGCGGCCTTTTTACGGTTCCTGGCCTTTTGCTG
GCCTTTTGCTCACATGTTCTTTCCTGCGTTATCCCCTGATTCTGTGGATAACCGTATTAC
CGCCTTTGAGTGAGCTGATACCGCTCGCCGCAGCCGAACGACCGAGCGCAGCGAGTCAGT
GAGCGAGGAAGCGGAAGAGCGCCCAATACGCAAACCGCCTCTCCCCGCGCGTTGGCCGAT
TCATTAATGCAGCTGGCACGACAGGTTTCCCGACTGGAAAGCGGGCAGTGAGCGCAACGC
AATTAATGTGAGTTAGCTCACTCATTAGGCACCCCAGGCTTTACACTTTATGCTTCCGGC
TCGTATGTTGTGTGGAATTGTGAGCGGATAACAATTTCACACAGGAAACAGCTATGACCA
TGATTACGCCAAGCTCGAAATTAACCCTCACTAAAGGGAACAAAAGCTGGTACCCCATAA
TAGTGTAGGTTGGGTTGTATATTATACAAAATAAATATTTTTTTTTGTCATAAAAAAAAT
GTTAACAAAACTTCAATCATTAAGTATTTAAATGATATGAAAAACAAAAAATAATATAAC
ATAATTGTTTTTTGTCTTATGCCTCATAAAATAAATAAAACAACTTTTAGTTCATGACCT
TTTCTAAATTTTTCTATTAAGGATATATTTATAATTATTGCTATTAAAATAAATACATAA
TTTTTATAGGTCTTTTTCAATAAAGATGCACTTTATTGGTGGTGAAAGCTCTAAAATTAT
AATAATGCTAATATTTTACTATATTAATAATTCTACAAAATAATAAAAAAGTTAAATAAA
GCATTTAAGATACTAAAAATTTATTGCCTATAATTAGATAAAACTATAAATTTTATTAAT
TCGAAACAATTTAAAAAATATCACATTAGCATACATAATTTAACAAATAATTAAAAAAAA
ATAATACATTTTATATTAAAATTTTTTTAATATTTTATTTGATATTTTTTTAAATATTTA
AATTATTATTATAAAACCCTAAAATAAAAATATCGCACAATAAATATTCATATTATGCAT
TTTTTATAAAACAAAATTTATTGACGAAATTAATTAAAAGAATATTTATTTTTTATATAA
GAATTATTAAATATAAAATTAAAAAATGAATACAAAACAGTCTAAGTATCGCAAAATTGT
GGCTTCATTCCTTTTGGCTGTTATATGCCTATTGCCGAAGCTTGCATGCCTGCAGGTCAA
CAATAAATAATAAATAAATATTGTGGAAATAAAATAACATATAATTATTTTTAATACATT
GATTTCCCTTTTATTTTTTTAAATTTCATTGATATAAAAATATATAATAATAACATATAT
GATTTCAAATTAATCTTTTCAAAAATGGTGTTTATTTTTGTATGTTTGTGTATGAATTAA
TCACATAACACATCTATTAAATTGAGTTGGTAATATAGACACAAATAAATATATATATTT
TTATAGCTTAAAAGTGTGTTATGAATATTTTAAGCATATTTTCTTTTTCTTTGGATTGTG
TAAAATGAACTCATATAATGCGTTTTTTTGTTTTTGTTATTTTGTCATTTTGTTATTTTG
CTATTTTATGGATTAATTTTTGTTTATAAAATGGGAAATAATTTTAACATATTTAAATAA
ATGGAGAAAAAATATAAAATAATTATAAAAAAAAGTTAATACACATTTTTTCCTGTTATA
GACCTTATATTTATTTATCCATATATATATATATATATATATATATATATATATACATAC
CAAGTGAATTAAGAGGAAAGCTAATTTATTATTCAGAATAATATATGAACTATATATAAT
TTTTATTATTTTGGTGTATATTAATCTGTCTATATGCATACATGCAATAATTTATCGACT
TATATATCAAATAACATAAAATAGAAGTGTTTTAAATTATGGATATATGCTCAATATTCA
TTTTTTTTAATAAGTTAGCTATATTTAAATTATACATTTTATATATGGTCTCTTTTTTTT
TTAAATATTATTTAAGTGATCATGAAAATATAAATAATTTTTTTTTATTTAATATCCTTT
TGCTTGCATGTGGTAAATGGAAATTTGGATGTGTTTTGAARGTTCGGATATAGTTGTATG
GACATATAATATATTTTGTGAAAAATTGGTTTTATGTTTATACTTATGCCAATACTTTTT
GAGTAAAACAAAGCAAGTGCTTATAAATAATTAAAGCCAATTTTATAATATATATTTTTT
TATTTAATTTGAATTTAGTAGTATAATTTTTTATGGTAAGTGCTCAAAGAGAGTTGCTTA
TAAAGTATGGTTTGTTTCTTTTTCGCCATTTTGAATTACACATTAAAAATATATAGATAC
ATATATTATAATATGAAATCATTAATAATTTAGGGAAATTCTACAAATTTAAAAACGAAT
AAAATAATTGTTTTTCATCATGCCATAACACAATATTGATATATACATGTACAAACATTT
TTTTTATTTGGAAAATATAAATTATATAAAAAAAAATGTATAGTATACAAAATGAGCATA
TTCACACGGGGTGGACGTTCATTTTTTCATTTTTCCCCTGTTTTTTATGAGTATATGATA
AAATTTTATGAACATTTACACAAAATGAAAATGGATATATAGGAAAAATGGAGCGGTATT
TCATTTATCTTTGATTGTCATTTGGATATTATATTACCYTGGGTAGGCAATTAAAAATGT
TAAATAACAATTTAAGGAAATTATATTTTATATATTAAAATTAACACTGTATTATATGAT
TCGCTTATAAAAGCCACTCTTTCCCCATGCAAAGCTGTTTAATATCAATTTTAACAAATT
ACACACATGTTAATATATTTATATATATAATTTATATATTTATAATTTATATATTTATAT
TTTTATTATTTATATATTTATTATTTATTGTGTGTGTCAATTCGGGTAGGATATACCTCT
TTTTTATTGTTTAAAGCGATTTGTATTCTAAAATATAAAGRATTTGAAAAAGAGAAAGAT
AGAATATGATCCCATCATATATAGCCCTATAATTTTTATTTAGCAGCGAATTAATTTTTC
TATTAAGTTTATGTGTAATTAAAATAACGGAATATATATAATACAATAAAAAAGTGCATA
AATTAAAATTTTTTCAATTAAATTTTTTTTTTTAAGGGGTTATATAATATTAAATATATA
AAATACGATTATATATTTTTGCTACAATTTTTTATATTAAGATATAAATAGTAAATAAAT
GGTATTATATGGCATGTAATATATAAATTTTTTCCAATTTTTATTTTATATACACTTTTC
CTTTTTTTGTCATAAAACTTAAACAATTTACACATTCATTTTAAAAATTGACTATTTGTT
TCAACATTTTTTGAGTTTCCGTTTTATAATAGTATTTTCATTTGTATATTGCTTATATAT
ATAAATACACACCTAAATGTTACAAAGGATCAATGCATAAACCGGTGTGTCTGGTCGTCG
CGATGACCCCCAAGAGGGGCATCGGCATCAACAACGGCCTCCCGTGGCCCCACTTGACCA
CAGATTTCAAACACTTTTCTCGTGTGACAAAAACGACGCCCGAAGAAGCCAGTCGCCTGA
ACGGGTGGCTTCCCAGGAAATTTGCAAAGACGGGCGACTCTGGACTTCCCTCTCCATCAG
TCGGCAAGAGATTCAACGCCGTTGTCATGGGACGGAAAACCTGGGAAAGCATGCCTCGAA
AGTTTAGACCCCTCGTGGACAGATTGAACATCGTCGTTTCCTCTTCCCTCAAAGAAGAAG
ACATTGCGGCGGAGAAGCCTCAAGCTGAAGGCCAGCAGCGCGTCCGAGTCTGTGCTTCAC
TCCCAGCAGCTCTCAGCCTTCTGGAGGAAGAGTACAAGGATTCTGTCGACCAGATTTTTG
TCGTGGGAGGAGCGGGACTGTACGAGGCAGCGCTGTCTCTGGGCGTTGCCTCTCACCTGT
ACATCACGCGTGTAGCCCGCGAGTTTCCGTGCGACGTTTTCTTCCCTGCGTTCCCCGGAG
ATGACATTCTTTCAAACAAATCAACTGCTGCGCAGGCTGCAGCTCCTGCCGAGTCTGTGT
TCGTTCCCTTTTGTCCGGAGCTCGGAAGAGAGAAGGACAATGAAGCGACGTATCGACCCA
TCTTCATTTCCAAGACCTTCTCAGACAACGGGGTTCCCTACGACTTTGTGGTTCTCGAGA
AGAGAAGGAAGACTGACGACGCAGCCACTGCGGAACCGAGCAACGCAATGAGCTCCTTGA
CGTCCACGAGGGAGACAACTCCCGTGCACGGGTTGCAGGCTCCTTCTTCGGCCGCAGCCA
TTGCCCCGGTGTTGGCGTGGATGGACGAAGAAGACCGGAAAAAACGCGAGCAAAAGGAAC
TGATTCGGGCCGTTCCGCATGTTCACTTTAGAGGCCATGAAGAGTTCCAGTACCTTGATC
TCATTGCCGACATTATTAACAATGGAAGGACAATGGATGACCGAACGGGCGTTGGTGTCA
TCTCCAAATTCGGCTGCACTATGCGCTACTCGCTGGATCAGGCCTTTCCACTTCTCACCA
CAAAGCGTGTGTTCTGGAAAGGGGTCCTCGAAGAGTTGCTGTGGTTCATTCGCGGCGACA
CGAACGCAAACCATCTTTCTGAGAAGGGCGTGAAGATCTGGGACAAGAATGTGACACGCG
AGTTCCTCGATTCGCGCAATCTCCCCCACCGAGAGGTCGGAGACATCGGCCCGGGCTACG
GCTTCCAGTGGAGACACTTCGGCGCGGCATACAAAGACATGCACACAGACTACACAGGGC
AGGGCGTCGACCAGCTGAAGAATGTGATCCAGATGCTGAGAACGAATCCAACAGATCGTC
GCATGCTCATGACTGCCTGGAATCCTGCAGCGCTGGACGAAATGGCGCTGCCGCCTTGTC
ACTTGTTGTGCCAGTTCTACGTGAACGACCAGAAGGAGCTGTCGTGCATCATGTATCAGC
GGTCGTGCGATGTCGGCCTCGGCGTCCCCTTCAACATCGCTTCCTATTCGCTTTTGACGC
TCATGGTTGCACACGTCTGCAACCTAAAACCTAAGGAGTTCATTCACTTCATGGGGAACA
CGCATGTCTACACGAACCATGTCGAGGCTTTAAAAGAGCAGCTGCGGAGAGAACCGAGAC
CGTTCCCCATTGTGAACATCCTCAACAAGGAACGCATCAAGGAAATCGACGATTTCACCG
CCGAGGATTTTGAGGTCGTGGGCTACGTCCCGCACGGACGAATCCAGATGGAGATGGCTG
TCTAGCGGAAATACAGAAGCTAGCTTTGATCCCGTTTTTCTTACTTATATATTTATACCA
ATTGATTGTATTTATAACTGTAAAAATGTGTATGTTGTGTGCATATTTTTTTTTGTGCAT
GCACATGCATGTAAATAGCTAAAATTATGAACATTTTATTTTTTGTTCAGAAAAAAAAAA
CTTTACACACATAAAATGGCTAGTATGAATAGCCATATTTTATATAAATTAAATCCTATG
AATTTATGACCATATTAAAAATTTAGATATTTATGGAACATAATATGTTTGAAACAATAA
GACAAAATTATTATTATTATTATTATTTTTACTGTTATAATTATGTTGTCTCTTCAATGA
TTCATAAATAGTTGGACTTGATTTTTAAAATGTTTATAATATGATTAGCATAGTTAAATA
AAAAAAGTTGAAAAATTAAAAAAAAACATATAAACACAAATGATGTTTTTTCCTTCAATT
TCGAT
Restriction sites to linearize plasmid KpnI/EcoRI
Partial or complete disruption of the geneUnknown
Additional remarks partial/complete disruption
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 1GCCGGTACCCCATAATAGTGTAGGTTGGGTTG
Additional information primer 1F1 primer
Sequence Primer 2GCCAAGCTTCGGCAATAGGCATATAACAGCC
Additional information primer 2R1 primer
Sequence Primer 3GCCGATATCCTTACTAAAGTGGTTCGCGACG
Additional information primer 3F2 primer
Sequence Primer 4GCCGAATTCCATAATAAGTTTTAATGCGTAAAC
Additional information primer 4R2 primer
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