Summary

RMgm-104
Malaria parasiteP. berghei
Genotype
Genetic modification not successful
DisruptedGene model (rodent): PBANKA_0209000; Gene model (P.falciparum): PF3D7_0104100; Gene product: conserved Plasmodium membrane protein, unknown function
PhenotypeNo phenotype has been described
Last modified: 18 February 2009, 11:43
  *RMgm-104
Successful modificationThe gene/parasite could not be changed/generated by the genetic modification.
The following genetic modifications were attempted Gene disruption
Number of attempts to introduce the genetic modification 3
Reference (PubMed-PMID number) Reference 1 (PMID number) : 18974882
Parent parasite used to introduce the genetic modification
Rodent Malaria ParasiteP. berghei
Parent strain/lineP. berghei ANKA
Name parent line/clone P. berghei ANKA 507cl1 (RMgm-7)
Other information parent lineP.berghei ANKA 507cl1 (RMgm-7) is a reference ANKA mutant line which expresses GFP under control of a constitutive promoter (PubMed: PMID: 16242190).
Attempts to generate the mutant parasite were performed by
Name PI/ResearcherE. Lasonder, C.J. Janse, H.G. Stunnenberg
Name Group/DepartmentDepartment of Molecular Biology
Name InstituteNCMLS, Radboud University Nijmegen
CityNijmegen
CountryThe Netherlands

  Disrupted: Mutant parasite with a disrupted gene
Details of the target gene
Gene Model of Rodent Parasite PBANKA_0209000
Gene Model P. falciparum ortholog PF3D7_0104100
Gene productconserved Plasmodium membrane protein, unknown function
Gene product: Alternative name
Details of the genetic modification
Inducable system usedNo
Additional remarks inducable system
Type of plasmid/construct usedPlasmid 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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CGATAAGCTTGCATGCCTGCAGGTCAACAATAAATAATAAATAAATATTGTGGAAATAAA
ATAACATATAATTATTTTTAATACATTGATTTCCCTTTTATTTTTTTAAATTTCATTGAT
ATAAAAATATATAATAATAACATATATGATTTCAAATTAATCTTTTCAAAAATGGTGTTT
ATTTTTGTATGTTTGTGTATGAATTAATCACATAACACATCTATTAAATTGAGTTGGTAA
TATAGACACAAATAAATATATATATTTTTATAGCTTAAAAGTGTGTTATGAATATTTTAA
GCATATTTTCTTTTTCTTTGGATTGTGTAAAATGAACTCATATAATGCGTTTTTTTGTTT
TTGTTATTTTGTCATTTTGTTATTTTGCTATTTTATGGATTAATTTTTGTTTATAAAATG
GGAAATAATTTTAACATATTTAAATAAATGGAGAAAAAATATAAAATAATTATAAAAAAA
AGTTAATACACATTTTTTCCTGTTATAGACCTTATATTTATTTATCCATATATATATATA
TATATATATATATATATATATACATACCAAGTGAATTAAGAGGAAAGCTAATTTATTATT
CAGAATAATATATGAACTATATATAATTTTTATTATTTTGGTGTATATTAATCTGTCTAT
ATGCATACATGCAATAATTTATCGACTTATATATCAAATAACATAAAATAGAAGTGTTTT
AAATTATGGATATATGCTCAATATTCATTTTTTTTAATAAGTTAGCTATATTTAAATTAT
ACATTTTATATATGGTCTCTTTTTTTTTTAAATATTATTTAAGTGATCATGAAAATATAA
ATAATTTTTTTTTATTTAATATCCTTTTGCTTGCATGTGGTAAATGGAAATTTGGATGTG
TTTTGAARGTTCGGATATAGTTGTATGGACATATAATATATTTTGTGAAAAATTGGTTTT
ATGTTTATACTTATGCCAATACTTTTTGAGTAAAACAAAGCAAGTGCTTATAAATAATTA
AAGCCAATTTTATAATATATATTTTTTTATTTAATTTGAATTTAGTAGTATAATTTTTTA
TGGTAAGTGCTCAAAGAGAGTTGCTTATAAAGTATGGTTTGTTTCTTTTTCGCCATTTTG
AATTACACATTAAAAATATATAGATACATATATTATAATATGAAATCATTAATAATTTAG
GGAAATTCTACAAATTTAAAAACGAATAAAATAATTGTTTTTCATCATGCCATAACACAA
TATTGATATATACATGTACAAACATTTTTTTTATTTGGAAAATATAAATTATATAAAAAA
AAATGTATAGTATACAAAATGAGCATATTCACACGGGGTGGACGTTCATTTTTTCATTTT
TCCCCTGTTTTTTATGAGTATATGATAAAATTTTATGAACATTTACACAAAATGAAAATG
GATATATAGGAAAAATGGAGCGGTATTTCATTTATCTTTGATTGTCATTTGGATATTATA
TTACCYTGGGTAGGCAATTAAAAATGTTAAATAACAATTTAAGGAAATTATATTTTATAT
ATTAAAATTAACACTGTATTATATGATTCGCTTATAAAAGCCACTCTTTCCCCATGCAAA
GCTGTTTAATATCAATTTTAACAAATTACACACATGTTAATATATTTATATATATAATTT
ATATATTTATAATTTATATATTTATATTTTTATTATTTATATATTTATTATTTATTGTGT
GTGTCAATTCGGGTAGGATATACCTCTTTTTTATTGTTTAAAGCGATTTGTATTCTAAAA
TATAAAGRATTTGAAAAAGAGAAAGATAGAATATGATCCCATCATATATAGCCCTATAAT
TTTTATTTAGCAGCGAATTAATTTTTCTATTAAGTTTATGTGTAATTAAAATAACGGAAT
ATATATAATACAATAAAAAAGTGCATAAATTAAAATTTTTTCAATTAAATTTTTTTTTTT
AAGGGGTTATATAATATTAAATATATAAAATACGATTATATATTTTTGCTACAATTTTTT
ATATTAAGATATAAATAGTAAATAAATGGTATTATATGGCATGTAATATATAAATTTTTT
CCAATTTTTATTTTATATACACTTTTCCTTTTTTTGTCATAAAACTTAAACAATTTACAC
ATTCATTTTAAAAATTGACTATTTGTTTCAACATTTTTTGAGTTTCCGTTTTATAATAGT
ATTTTCATTTGTATATTGCTTATATATATAAATACACACCTAAATGTTACAAAGGATCAA
TGCATAAACCGGTGTGTCTGGTCGTCGCGATGACCCCCAAGAGGGGCATCGGCATCAACA
ACGGCCTCCCGTGGCCCCACTTGACCACAGATTTCAAACACTTTTCTCGTGTGACAAAAA
CGACGCCCGAAGAAGCCAGTCGCCTGAACGGGTGGCTTCCCAGGAAATTTGCAAAGACGG
GCGACTCTGGACTTCCCTCTCCATCAGTCGGCAAGAGATTCAACGCCGTTGTCATGGGAC
GGAAAACCTGGGAAAGCATGCCTCGAAAGTTTAGACCCCTCGTGGACAGATTGAACATCG
TCGTTTCCTCTTCCCTCAAAGAAGAAGACATTGCGGCGGAGAAGCCTCAAGCTGAAGGCC
AGCAGCGCGTCCGAGTCTGTGCTTCACTCCCAGCAGCTCTCAGCCTTCTGGAGGAAGAGT
ACAAGGATTCTGTCGACCAGATTTTTGTCGTGGGAGGAGCGGGACTGTACGAGGCAGCGC
TGTCTCTGGGCGTTGCCTCTCACCTGTACATCACGCGTGTAGCCCGCGAGTTTCCGTGCG
ACGTTTTCTTCCCTGCGTTCCCCGGAGATGACATTCTTTCAAACAAATCAACTGCTGCGC
AGGCTGCAGCTCCTGCCGAGTCTGTGTTCGTTCCCTTTTGTCCGGAGCTCGGAAGAGAGA
AGGACAATGAAGCGACGTATCGACCCATCTTCATTTCCAAGACCTTCTCAGACAACGGGG
TTCCCTACGACTTTGTGGTTCTCGAGAAGAGAAGGAAGACTGACGACGCAGCCACTGCGG
AACCGAGCAACGCAATGAGCTCCTTGACGTCCACGAGGGAGACAACTCCCGTGCACGGGT
TGCAGGCTCCTTCTTCGGCCGCAGCCATTGCCCCGGTGTTGGCGTGGATGGACGAAGAAG
ACCGGAAAAAACGCGAGCAAAAGGAACTGATTCGGGCCGTTCCGCATGTTCACTTTAGAG
GCCATGAAGAGTTCCAGTACCTTGATCTCATTGCCGACATTATTAACAATGGAAGGACAA
TGGATGACCGAACGGGCGTTGGTGTCATCTCCAAATTCGGCTGCACTATGCGCTACTCGC
TGGATCAGGCCTTTCCACTTCTCACCACAAAGCGTGTGTTCTGGAAAGGGGTCCTCGAAG
AGTTGCTGTGGTTCATTCGCGGCGACACGAACGCAAACCATCTTTCTGAGAAGGGCGTGA
AGATCTGGGACAAGAATGTGACACGCGAGTTCCTCGATTCGCGCAATCTCCCCCACCGAG
AGGTCGGAGACATCGGCCCGGGCTACGGCTTCCAGTGGAGACACTTCGGCGCGGCATACA
AAGACATGCACACAGACTACACAGGGCAGGGCGTCGACCAGCTGAAGAATGTGATCCAGA
TGCTGAGAACGAATCCAACAGATCGTCGCATGCTCATGACTGCCTGGAATCCTGCAGCGC
TGGACGAAATGGCGCTGCCGCCTTGTCACTTGTTGTGCCAGTTCTACGTGAACGACCAGA
AGGAGCTGTCGTGCATCATGTATCAGCGGTCGTGCGATGTCGGCCTCGGCGTCCCCTTCA
ACATCGCTTCCTATTCGCTTTTGACGCTCATGGTTGCACACGTCTGCAACCTAAAACCTA
AGGAGTTCATTCACTTCATGGGGAACACGCATGTCTACACGAACCATGTCGAGGCTTTAA
AAGAGCAGCTGCGGAGAGAACCGAGACCGTTCCCCATTGTGAACATCCTCAACAAGGAAC
GCATCAAGGAAATCGACGATTTCACCGCCGAGGATTTTGAGGTCGTGGGCTACGTCCCGC
ACGGACGAATCCAGATGGAGATGGCTGTCTAGCGGAAATACAGAAGCTAGCTTTGATCCC
GTTTTTCTTACTTATATATTTATACCAATTGATTGTATTTATAACTGTAAAAATGTGTAT
GTTGTGTGCATATTTTTTTTTGTGCATGCACATGCATGTAAATAGCTAAAATTATGAACA
TTTTATTTTTTGTTCAGAAAAAAAAAACTTTACACACATAAAATGGCTAGTATGAATAGC
CATATTTTATATAAATTAAATCCTATGAATTTATGACCATATTAAAAATTTAGATATTTA
TGGAACATAATATGTTTGAAACAATAAGACAAAATTATTATTATTATTATTATTTTTACT
GTTATAATTATGTTGTCTCTTCAATGATTCATAAATAGTTGGACTTGATTTTTAAAATGT
TTATAATATGATTAGCATAGTTAAATAAAAAAAGTTGAAAAATTAAAAAAAAACATATAA
ACACAAATGATGTTTTTTCCTTCAATTTCGATATCATTATAAATGAAAATATGTGGATAA
TTATTCCTACAGATAATAATAAATATGTAAAAAATGTTAAATCAGATATTATTAAAAAAT
CATTATTAGGAACATGTTTAACAAAAGAAAGTGCCCAATTTGAAGAGTATCATCTTATGG
GAACAGATGCTTATATGAAATATATAAATAAATTTGGTTTGCTAAATAATTATGAGATGT
GTTTTGAAGACCCATCATGTGAAAATAACGACAGAAAATACAATATCAATTATAACTCTA
AAGTTACAGACCCAAAATATCTTGATGTTAAACGTAATAGAGTCATGCTTTATCAAGATA
GCGATTTTGATAATGTACTTGAAGTGTTCATATTAAAATCAAAAATTAATAATGATAAAA
TTTTTAATATTAGCGATTTAGATGAAACAAAGAAGGAAAATATAACATGGAGAGAATATA
CACCAAAAAATGGAGCCGGAGAAAATAAAAAATCTATTGTTCAAACATATTTTGAGAAAG
CTATTGAATATATGAAATTTGAAAATGTTTTAACTTTACTTAAAGAAGTTAATAATCATA
TAAATTCATTTAAAAATGTTATTATTGAAAAAGCTAATTCATTAGTAGATAATACAAATT
GTAGTAGATTTATTAATGTACTAACTAATATAAGACATAATTATTGTGACAATGGAATTT
TAAAATTAACTCGATTATCAGTCATACTTATTTCATGTGGATTTGTTTCCTTTTGTCTTT
GGTACCTTTTCCTTTTTTTTTGGATATACCATCAAATGAAGATTATTTGAATTTATTTTT
AACTCATATTAATTTTCATCGAAAAATAAAATTTACTTTTTCACCATAAAAAATATTGGA
AATCTCCAATATTTTTTAAATAAAACCCGACAGATCTTGTCTTTTATATATTTCTATCAT
ATAATGTTATATTTTTTTGCCCCAATTTGTAATAATAATCCGGGATCCACTAGTTCTAGA
GCGGCCGCCACCGCGGTGGAGCTCCAATTCGCCCTATAGTGAGTCGTATTACGCGCGCTC
ACTGGCCGTCGTTTTACAACGTCGTGACTGGGAAAACCCTGGCGTTACCCAACTTAATCG
CCTTGCAGCACATCCCCCTTTCGCCAGCTGGCGTAATAGCGAAGAGGCCCGCACCGATCG
CCCTTCCCAACAGTTGCGCAGCCTGAATGGCGAATGGGACGCGCCCTGTAGCGGCGCATT
AAGCGCGGCGGGTGTGGTGGTTACGCGCAGCGTGACCGCTACACTTGCCAGCGCCCTAGC
GCCCGCTCCTTTCGCTTTCTTCCCTTCCTTTCTCGCCACGTTCGCCGGCTTTCCCCGTCA
AGCTCTAAATCGGGGGCTCCCTTTAGGGTTCCGATTTAGTGCTTTACGGCACCTCGACCC
CAAAAAACTTGATTAGGGTGATGGTTCACGTAGTGGGCCATCGCCCTGATAGACGGTTTT
TCGCCCTTTGACGTTGGAGTCCACGTTCTTTAATAGTGGACTCTTGTTCCAAACTGGAAC
AACACTCAACCCTATCTCGGTCTATTCTTTTGATTTATAAGGGATTTTGCCGATTTCGGC
CTATTGGTTAAAAAATGAGCTGATTTAACAAAAATTTAACGCGAATTTTAACAAAATATT
AACGCTTACAATTTAGGTGGCACTTTTCGGGGAAATGTGCGCGGAACCCCTATTTGTTTA
TTTTTCTAAATACATTCAAATATGTATCCGCTCATGAGACAATAACCCTGATAAATGCTT
CAATAATATTGAAAAAGGAAGAGTATGAGTATTCAACATTTCCGTGTCGCCCTTATTCCC
TTTTTTGCGGCATTTTGCCTTCCTGTTTTTGCTCACCCAGAAACGCTGGTGAAAGTAAAA
GATGCTGAAGATCAGTTGGGTGCACGAGTGGGTTACATCGAACTGGATCTCAACAGCGGT
AAGATCCTTGAGAGTTTTCGCCCCGAAGAACGTTTTCCAATGATGAGCACTTTTAAAGTT
CTGCTATGTGGCGCGGTATTATCCCGTATTGACGCCGGGCAAGAGCAACTCGGTCGCCGC
ATACACTATTCTCAGAATGACTTGGTTGAGTACTCACCAGTCACAGAAAAGCATCTTACG
GATGGCATGACAGTAAGAGAATTATGCAGTGCTGCCATAACCATGAGTGATAACACTGCG
GCCAACTTACTTCTGACAACGATCGGAGGACCGAAGGAGCTAACCGCTTTTTTGCACAAC
ATGGGGGATCATGTAACTCGCCTTGATCGTTGGGAACCGGAGCTGAATGAAGCCATACCA
AACGACGAGCGTGACACCACGATGCCTGTAGCAATGGCAACAACGTTGCGCAAACTATTA
ACTGGCGAACTACTTACTCTAGCTTCCCGGCAACAATTAATAGACTGGATGGAGGCGGAT
AAAGTTGCAGGACCACTTCTGCGCTCGGCCCTTCCGGCTGGCTGGTTTATTGCTGATAAA
TCTGGAGCCGGTGAGCGTGGGTCTCGCGGTATCATTGCAGCACTGGGGCCAGATGGTAAG
CCCTCCCGTATCGTAGTTATCTACACGACGGGGAGTCAGGCAACTATGGATGAACGAAAT
AGACAGATCGCTGAGATAGGTGCCTCACTGATTAAGCATTGGTAACTGTCAGACCAAGTT
TACTCATATATACTTTAGATTGATTTAAAACTTCATTTTTAATTTAAAAGGATCTAGGTG
AAGATCCTTTTTGATAATCTCATGACCAAAATCCCTTAACGTGAGTTTTCGTTCCACTGA
GCGTCAGACCCCGTAGAAAAGATCAAAGGATCTTCTTGAGATCCTTTTTTTCTGCGCGTA
ATCTGCTGCTTGCAAACAAAAAAACCACCGCTACCAGCGGTGGTTTGTTTGCCGGATCAA
GAGCTACCAACTCTTTTTCCGAAGGTAACTGGCTTCAGCAGAGCGCAGATACCAAATACT
GTCCTTCTAGTGTAGCCGTAGTTAGGCCACCACTTCAAGAACTCTGTAGCACCGCCTACA
TACCTCGCTCTGCTAATCCTGTTACCAGTGGCTGCTGCCAGTGGCGATAAGTCGTGTCTT
ACCGGGTTGGACTCAAGACGATAGTTACCGGATAAGGCGCAGCGGTCGGGCTGAACGGGG
GGTTCGTGCACACAGCCCAGCTTGGAGCGAACGACCTACACCGAACTGAGATACCTACAG
CGTGAGCTATGAGAAAGCGCCACGCTTCCCGAAGGGAGAAAGGCGGACAGGTATCCGGTA
AGCGGCAGGGTCGGAACAGGAGAGCGCACGAGGGAGCTTCCAGGGGGAAACGCCTGGTAT
CTTTATAGTCCTGTCGGGTTTCGCCACCTCTGACTTGAGCGTCGATTTTTGTGATGCTCG
TCAGGGGGGCGGAGCCTATGGAAAAACGCCAGCAACGCGGCCTTTTTACGGTTCCTGGCC
TTTTGCTGGCCTTTTGCTCACATGTTCTTTCCTGCGTTATCCCCTGATTCTGTGGATAAC
CGTATTACCGCCTTTGAGTGAGCTGATACCGCTCGCCGCAGCCGAACGACCGAGCGCAGC
GAGTCAGTGAGCGAGGAAGCGGAAGAGCGCCCAATACGCAAACCGCCTCTCCCCGCGCGT
TGGCCGATTCATTAATGCAGCTGGCACGACAGGTTTCCCGACTGGAAAGCGGGCAGTGAG
CGCAACGCAATTAATGTGAGTTAGCTCACTCATTAGGCACCCCAGGCTTTACACTTTATG
CTTCCGGCTCGTATGTTGTGTGGAATTGTGAGCGGATAACAATTTCACACAGGAAACAGC
TATGACCATGATTACGCCAAGCTCGAAATTAACCCTCACTAAAGGGAACAAAAGCTGGTA
CCCGTTTATATTCATCATGGAGAATGGAAGATCTTAATAATTTTAACGGGAAAAGTGTTA
TAAAAATAATGGAAAGAAATCCATATGTTATATTTTTTTTTTTTTTTATAATGATTTTTA
TTATTGTTTATTTAATTTCATTTATTTTGTATACTAAATGGTTTAAAAAATTATTAAAAA
AATTTTCGAATTCACACAAAAATAATAAAGATAAAGAAGAAGATTGGGTAAAAAAAAATA
AAGCTTATAGAAATTCTAATAGCACACATGGTACTATTAATAAGGATAATTATAATCAGG
AACTTGATGAGCTTCATAATAGTGATGAAAATGAAGAAAATAGTAATGTTATAAATATTG
TAAAAAAGAGAGCTTATAATTTAGTAATTAATTTGATAGTTTGTTCTTTTCTTATTTGTC
TTATTTTTTTGGGAATTTGGACAATATTTATTTTTGCAGATACACAAAAAGGAATTAATA
TGAATATCTGTGGATTATCAAAAACAGTTGAACAGTTTCTTATTGATAAATGCCCCGACA
CAAAAAATGTAAATCCACAATGTTATTCTTTAGAGCATGTTATTAATGATGCTGTTTCAG
TAATGAATCAGTATCAACTCACCAAAGAATTTGTTAAAAATAAAACGAATTTGAACAAGA
ATAAGGGCTTGCCTATTGTTTTAAGTAGGTTTATATCACTACCCGCGTTTCTCCTTTGCA
CCTAAACAAGTTGCGTCTGTTTTTGCATCTAAAATTTGCGGCTGCTCCTTCACATCTAAA
AATGTTGCGTCTGCTTTTGCGTCTGCTTTTGCGTCTGCTTTTGCATCTGCCCCTTCACAT
CTATTTCTTTTTTCCCCGTATAGAGTACCAAACCGGATTCAACATGCTAGCAAAACTTAG
AGAAT
Restriction sites to linearize plasmid
Partial or complete disruption of the genePartial
Additional remarks partial/complete disruption See for more information about the disruption Lasonder, E. et al., 2008, PloS Pathogens 10, e10000195 (Supporting Information: S3).
Selectable marker used to select the mutant parasitetgdhfr
Promoter of the selectable markerpbdhfr
Selection (positive) procedurepyrimethamine
Selection (negative) procedureNo
Additional remarks genetic modificationThe protein was detected in a proteome analysis of P. falciparum oocysts and sporozoites and was found in oocyst-derived sporozoites and in salivary gland sporozoites. It contains a transmembrane region.
See for more information about the disruption Lasonder, E. et al., 2008, PloS Pathogens 10, e10000195 (Supporting Information: S3).
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 1CGGGGTACCCGTTTATATTCATCATGGAGAATGG
Additional information primer 1L2636 (Asp718); 5'
Sequence Primer 2CCATCGATTCTCTAAGTTTTGCTAGC
Additional information primer 2L2637 (ClaI); 5'
Sequence Primer 3CCGGATATCATTATAAATGAAAATATGTGGATAATTATTCC
Additional information primer 3L2638 (EcoRV); 3'
Sequence Primer 4CGCGGATCCCGGATTATTATTACAAATTGGGG
Additional information primer 4L2639 (BamHI); 3'
Sequence Primer 5
Additional information primer 5
Sequence Primer 6
Additional information primer 6