Summary

RMgm-103
Malaria parasiteP. berghei
Genotype
Genetic modification not successful
DisruptedGene model (rodent): PBANKA_1327100; Gene model (P.falciparum): PF3D7_1463900; Gene product: conserved Plasmodium membrane protein, unknown function
PhenotypeNo phenotype has been described
Last modified: 6 March 2009, 21:27
  *RMgm-103
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_1327100
Gene Model P. falciparum ortholog PF3D7_1463900
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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GCGGATAACAATTTCACACAGGAAACAGCTATGACCATGATTACGCCAAGCTCGAAATTA
ACCCTCACTAAAGGGAACAAAAGCTGGTACCTTCACTAGCGTAATTCTGAAACAACGCAT
TAAAAAGACAATTCATTTTTACTTCTACTAATATATCTGTCTTATTATTATTATATGCAT
TTTTCTCGATATTTTTTTTCATTTATATTTATTTATTTATTTATTTATTTTTCTTTTTTT
GAAGTTTGTAGAAAAGAAAAAGAAATACACAACACAGCATGTAATTTTTAGGACGTTTCT
AAAAAAACCATATTAATAAATGGTAACTAGTTTTAAAATTACAAAATCATCTCATTTTTA
AGGTTAGAATAAAATTTATTATTTTTGTTATATGTTATAACATTTTGATTATAATTATAC
TATTATGGTTTTGTGAAAAATGAAATAAAGTGGATTTTTAAAATTATAAAAACATGTATA
CATTATATAAAAAACGAATCCATATTATTTAAATGAAAAATCGAAAATAACAATAAAATC
ATTTTTTTTTTTCATTTTATTAGCAGGACCTATAATATGTTAAAACTCTATCCTCTAATT
TTTTTATGTGTTCGTTAAAGAAAATAGTGAAAAGAAATATATTATTTTCTATATAGATAT
AATGGATTTATCATTTATGAATGAAAACATATATATTTATGCACATCCATAAATTAATAT
TTATAATAAACGAATGAAACGTTTTCATACTTGTACATTAAAAATTAATAATACAAACAT
GCAAATGTGTTTTTATTCAATTGTATACGATATTCTTCGAAAATCATCATAGCAAGCTTG
CATGCCTGCAGGTCAACAATAAATAATAAATAAATATTGTGGAAATAAAATAACATATAA
TTATTTTTAATACATTGATTTCCCTTTTATTTTTTTAAATTTCATTGATATAAAAATATA
TAATAATAACATATATGATTTCAAATTAATCTTTTCAAAAATGGTGTTTATTTTTGTATG
TTTGTGTATGAATTAATCACATAACACATCTATTAAATTGAGTTGGTAATATAGACACAA
ATAAATATATATATTTTTATAGCTTAAAAGTGTGTTATGAATATTTTAAGCATATTTTCT
TTTTCTTTGGATTGTGTAAAATGAACTCATATAATGCGTTTTTTTGTTTTTGTTATTTTG
TCATTTTGTTATTTTGCTATTTTATGGATTAATTTTTGTTTATAAAATGGGAAATAATTT
TAACATATTTAAATAAATGGAGAAAAAATATAAAATAATTATAAAAAAAAGTTAATACAC
ATTTTTTCCTGTTATAGACCTTATATTTATTTATCCATATATATATATATATATATATAT
ATATATATATACATACCAAGTGAATTAAGAGGAAAGCTAATTTATTATTCAGAATAATAT
ATGAACTATATATAATTTTTATTATTTTGGTGTATATTAATCTGTCTATATGCATACATG
CAATAATTTATCGACTTATATATCAAATAACATAAAATAGAAGTGTTTTAAATTATGGAT
ATATGCTCAATATTCATTTTTTTTAATAAGTTAGCTATATTTAAATTATACATTTTATAT
ATGGTCTCTTTTTTTTTTAAATATTATTTAAGTGATCATGAAAATATAAATAATTTTTTT
TTATTTAATATCCTTTTGCTTGCATGTGGTAAATGGAAATTTGGATGTGTTTTGAARGTT
CGGATATAGTTGTATGGACATATAATATATTTTGTGAAAAATTGGTTTTATGTTTATACT
TATGCCAATACTTTTTGAGTAAAACAAAGCAAGTGCTTATAAATAATTAAAGCCAATTTT
ATAATATATATTTTTTTATTTAATTTGAATTTAGTAGTATAATTTTTTATGGTAAGTGCT
CAAAGAGAGTTGCTTATAAAGTATGGTTTGTTTCTTTTTCGCCATTTTGAATTACACATT
AAAAATATATAGATACATATATTATAATATGAAATCATTAATAATTTAGGGAAATTCTAC
AAATTTAAAAACGAATAAAATAATTGTTTTTCATCATGCCATAACACAATATTGATATAT
ACATGTACAAACATTTTTTTTATTTGGAAAATATAAATTATATAAAAAAAAATGTATAGT
ATACAAAATGAGCATATTCACACGGGGTGGACGTTCATTTTTTCATTTTTCCCCTGTTTT
TTATGAGTATATGATAAAATTTTATGAACATTTACACAAAATGAAAATGGATATATAGGA
AAAATGGAGCGGTATTTCATTTATCTTTGATTGTCATTTGGATATTATATTACCYTGGGT
AGGCAATTAAAAATGTTAAATAACAATTTAAGGAAATTATATTTTATATATTAAAATTAA
CACTGTATTATATGATTCGCTTATAAAAGCCACTCTTTCCCCATGCAAAGCTGTTTAATA
TCAATTTTAACAAATTACACACATGTTAATATATTTATATATATAATTTATATATTTATA
ATTTATATATTTATATTTTTATTATTTATATATTTATTATTTATTGTGTGTGTCAATTCG
GGTAGGATATACCTCTTTTTTATTGTTTAAAGCGATTTGTATTCTAAAATATAAAGRATT
TGAAAAAGAGAAAGATAGAATATGATCCCATCATATATAGCCCTATAATTTTTATTTAGC
AGCGAATTAATTTTTCTATTAAGTTTATGTGTAATTAAAATAACGGAATATATATAATAC
AATAAAAAAGTGCATAAATTAAAATTTTTTCAATTAAATTTTTTTTTTTAAGGGGTTATA
TAATATTAAATATATAAAATACGATTATATATTTTTGCTACAATTTTTTATATTAAGATA
TAAATAGTAAATAAATGGTATTATATGGCATGTAATATATAAATTTTTTCCAATTTTTAT
TTTATATACACTTTTCCTTTTTTTGTCATAAAACTTAAACAATTTACACATTCATTTTAA
AAATTGACTATTTGTTTCAACATTTTTTGAGTTTCCGTTTTATAATAGTATTTTCATTTG
TATATTGCTTATATATATAAATACACACCTAAATGTTACAAAGGATCAATGCATAAACCG
GTGTGTCTGGTCGTCGCGATGACCCCCAAGAGGGGCATCGGCATCAACAACGGCCTCCCG
TGGCCCCACTTGACCACAGATTTCAAACACTTTTCTCGTGTGACAAAAACGACGCCCGAA
GAAGCCAGTCGCCTGAACGGGTGGCTTCCCAGGAAATTTGCAAAGACGGGCGACTCTGGA
CTTCCCTCTCCATCAGTCGGCAAGAGATTCAACGCCGTTGTCATGGGACGGAAAACCTGG
GAAAGCATGCCTCGAAAGTTTAGACCCCTCGTGGACAGATTGAACATCGTCGTTTCCTCT
TCCCTCAAAGAAGAAGACATTGCGGCGGAGAAGCCTCAAGCTGAAGGCCAGCAGCGCGTC
CGAGTCTGTGCTTCACTCCCAGCAGCTCTCAGCCTTCTGGAGGAAGAGTACAAGGATTCT
GTCGACCAGATTTTTGTCGTGGGAGGAGCGGGACTGTACGAGGCAGCGCTGTCTCTGGGC
GTTGCCTCTCACCTGTACATCACGCGTGTAGCCCGCGAGTTTCCGTGCGACGTTTTCTTC
CCTGCGTTCCCCGGAGATGACATTCTTTCAAACAAATCAACTGCTGCGCAGGCTGCAGCT
CCTGCCGAGTCTGTGTTCGTTCCCTTTTGTCCGGAGCTCGGAAGAGAGAAGGACAATGAA
GCGACGTATCGACCCATCTTCATTTCCAAGACCTTCTCAGACAACGGGGTTCCCTACGAC
TTTGTGGTTCTCGAGAAGAGAAGGAAGACTGACGACGCAGCCACTGCGGAACCGAGCAAC
GCAATGAGCTCCTTGACGTCCACGAGGGAGACAACTCCCGTGCACGGGTTGCAGGCTCCT
TCTTCGGCCGCAGCCATTGCCCCGGTGTTGGCGTGGATGGACGAAGAAGACCGGAAAAAA
CGCGAGCAAAAGGAACTGATTCGGGCCGTTCCGCATGTTCACTTTAGAGGCCATGAAGAG
TTCCAGTACCTTGATCTCATTGCCGACATTATTAACAATGGAAGGACAATGGATGACCGA
ACGGGCGTTGGTGTCATCTCCAAATTCGGCTGCACTATGCGCTACTCGCTGGATCAGGCC
TTTCCACTTCTCACCACAAAGCGTGTGTTCTGGAAAGGGGTCCTCGAAGAGTTGCTGTGG
TTCATTCGCGGCGACACGAACGCAAACCATCTTTCTGAGAAGGGCGTGAAGATCTGGGAC
AAGAATGTGACACGCGAGTTCCTCGATTCGCGCAATCTCCCCCACCGAGAGGTCGGAGAC
ATCGGCCCGGGCTACGGCTTCCAGTGGAGACACTTCGGCGCGGCATACAAAGACATGCAC
ACAGACTACACAGGGCAGGGCGTCGACCAGCTGAAGAATGTGATCCAGATGCTGAGAACG
AATCCAACAGATCGTCGCATGCTCATGACTGCCTGGAATCCTGCAGCGCTGGACGAAATG
GCGCTGCCGCCTTGTCACTTGTTGTGCCAGTTCTACGTGAACGACCAGAAGGAGCTGTCG
TGCATCATGTATCAGCGGTCGTGCGATGTCGGCCTCGGCGTCCCCTTCAACATCGCTTCC
TATTCGCTTTTGACGCTCATGGTTGCACACGTCTGCAACCTAAAACCTAAGGAGTTCATT
CACTTCATGGGGAACACGCATGTCTACACGAACCATGTCGAGGCTTTAAAAGAGCAGCTG
CGGAGAGAACCGAGACCGTTCCCCATTGTGAACATCCTCAACAAGGAACGCATCAAGGAA
ATCGACGATTTCACCGCCGAGGATTTTGAGGTCGTGGGCTACGTCCCGCACGGACGAATC
CAGATGGAGATGGCTGTCTAGCGGAAATACAGAAGCTAGCTTTGATCCCGTTTTTCTTAC
TTATATATTTATACCAATTGATTGTATTTATAACTGTAAAAATGTGTATGTTGTGTGCAT
ATTTTTTTTTGTGCATGCACATGCATGTAAATAGCTAAAATTATGAACATTTTATTTTTT
GTTCAGAAAAAAAAAACTTTACACACATAAAATGGCTAGTATGAATAGCCATATTTTATA
TAAATTAAATCCTATGAATTTATGACCATATTAAAAATTTAGATATTTATGGAACATAAT
ATGTTTGAAACAATAAGACAAAATTATTATTATTATTATTATTTTTACTGTTATAATTAT
GTTGTCTCTTCAATGATTCATAAATAGTTGGACTTGATTTTTAAAATGTTTATAATATGA
TTAGCATAGTTAAATAAAAAAAGTTGAAAAATTAAAAAAAAACATATAAACACAAATGAT
GTTTTTTCCTTCAATTTCGATATCGAATTCGTGAATTATATTAACAGCATTTATAAATAA
ATCACTATATGAGTATTACACGACGTTCTATTACATATGTATTTTCTTAAAATAAATAAG
CAACATCTGTAGAGATAATACAGCATTATATAGAGAAATATTTATGAGTCTAATAGTTCC
TTTCTTTTTCTTTTATTATATTAATACATGCATGAATATATACTTTTTTTTCAGAACATG
TCAGATACAAAATAAATATAGGATGCCAATTGATTTACTTATATTCCATATTTGATTTTT
AATATATATTTTTAAAAGGAGTTTCCTATTAATTTAAGAAAACATTAGTGTAAAGAAAAA
TGATAGAATTTCTCATTGTTTTTACAATTTTCTTTTTTTATGTTTATTGTTTTCGTTTTT
ATTTATTTATTTAGTTATTTTTTAGGGGTTGCTATTATTTTTATTTTATCGTTTATTTTA
TTATTACATTTACGACAATGTAATTTTTATGTGCTAATTTTTTGTTATTTTAACTATCAC
TTCTCTTTATATTTTCTTTTTGAAAGTATGCTGAATATACGCGTTATATATTTTTTTTCA
TTAAACTTTAGGCTTAATTTTTTATTTTTATAATTATGTTGTAATTAATATTTTATTTTC
CTTATTAGTATTTTTAAAATAATAGTTTTTTAATCTCTGTTTTTTAAAATTTTATATGAT
TTCTTTATATATGCATTTTTGTTCGATTCCATTCATGTTAATGTGGATGAATTTATTTTA
TATAAGAAACTGGCACTTATTTACCTTTTTAATTTTAATATTAAATAAAGTCCAATACAA
AAATATAAAATGGTGTGTATTTTATAATATGTATTTAGTATAGTGAGCAAATGTATATTT
ATATTCTTAATGCATAAAGAAAAGGAATATTGTAGAATATAAACAGTAAATGGTAACACA
TATGTTAATGCGGCCGCCACCGCGGTGGAGCTCCAATTCGCCCTATAGTGAGTCGTATTA
CGCGCGCTCACTGGCCGTCGTTTTACAACGTCGTGACTGGGAAAACCCTGGCGTTACCCA
ACTTAATCGCCTTGCAGCACATCCCCCTTTCGCCAGCTGGCGTAATAGCGAAGAGGCCCG
CACCGATCGCCCTTCCCAACAGTTGCGCAGCCTGAATGGCGAATGGGACGCGCCCTGTAG
CGGCGCATTAAGCGCGGCGGGTGTGGTGGTTACGCGCAGCGTGACCGCTACACTTGCCAG
CGCCCTAGCGCCCGCTCCTTTCGCTTTCTTCCCTTCCTTTCTCGCCACGTTCGCCGGCTT
TCCCCGTCAAGCTCTAAATCGGGGGCTCCCTTTAGGGTTCCGATTTAGTGCTTTACGGCA
CCTCGACCCCAAAAAACTTGATTAGGGTGATGGTTCACGTAGTGGGCCATCGCCCTGATA
GACGGTTTTTCGCCCTTTGACGTTGGAGTCCACGTTCTTTAATAGTGGACTCTTGTTCCA
AACTGGAACAACACTCAACCCTATCTCGGTCTATTCTTTTGATTTATAAGGGATTTTGCC
GATTTCGGCCTATTGGTTAAAAAATGAGCTGATTTAACAAAAATTTAACGCGAATTTTAA
CAAAATATTAACGCTTACAATTTAGGTGGCACTTTTCGGGGAAATGTGCGCGGAACCCCT
ATTTGTTTATTTTTCTAAATACATTCAAATATGTATCCGCTCATGAGACAATAACCCTGA
TAAATGCTTCAATAATATTGAAAAAGGAAGAGTATGAGTATTCAACATTTCCGTGTCGCC
CTTATTCCCTTTTTTGCGGCATTTTGCCTTCCTGTTTTTGCTCACCCAGAAACGCTGGTG
AAAGTAAAAGATGCTGAAGATCAGTTGGGTGCACGAGTGGGTTACATCGAACTGGATCTC
AACAGCGGTAAGATCCTTGAGAGTTTTCGCCCCGAAGAACGTTTTCCAATGATGAGCACT
TTTAAAGTTCTGCTATGTGGCGCGGTATTATCCCGTATTGACGCCGGGCAAGAGCAACTC
GGTCGCCGCATACACTATTCTCAGAATGACTTGGTTGAGTACTCACCAGTCACAGAAAAG
CATCTTACGGATGGCATGACAGTAAGAGAATTATGCAGTGCTGCCATAACCATGAGTGAT
AACACTGCGGCCAACTTACTTCTGACAACGATCGGAGGACCGAAGGAGCTAACCGCTTTT
TTGCACAACATGGGGGATCATGTAACTCGCCTTGATCGTTGGGAACCGGAGCTGAATGAA
GCCATACCAAACGACGAGCGTGACACCACGATGCCTGTAGCAATGGCAACAACGTTGCGC
AAACTATTAACTGGCGAACTACTTACTCTAGCTTCCCGGCAACAATTAATAGACTGGATG
GAGGCGGATAAAGTTGCAGGACCACTTCTGCGCTCGGCCCTTCCGGCTGGCTGGTTTATT
GCTGATAAATCTGGAGCCGGTGAGCGTGGGTCTCGCGGTATCATTGCAGCACTGGGGCCA
GATGGTAAGCCCTCCCGTATCGTAGTTATCTACACGACGGGGAGTCAGGCAACTATGGAT
GAACGAAATAGACAGATCGCTGAGATAGGTGCCTCACTGATTAAGCATTGGTAACTGTCA
GACCAAGTTTACTCATATATACTTTAGATTGATTTAAAACTTCATTTTTAATTTAAAAGG
ATCTAGGTGAAGATCCTTTTTGATAATCTCATGACCAAAATCCCTTAACGTGAGTTTTCG
TTCCACTGAGCGTCAGACCCCGTAGAAAAGATCAAAGGATCTTCTTGAGATCCTTTTTTT
CTGCGCGTAATCTGCTGCTTGCAAACAAAAAAACCACCGCTACCAGCGGTGGTTTGTTTG
CCGGATCAAGAGCTACCAACTCTTTTTCCGAAGGTAACTGGCTTCAGCAGAGCGCAGATA
CCAAATACTGTCCTTCTAGTGTAGCCGTAGTTAGGCCACCACTTCAAGAACTCTGTAGCA
CCGCCTACATACCTCGCTCTGCTAATCCTGTTACCAGTGGCTGCTGCCAGTGGCGATAAG
TCGTGTCTTACCGGGTTGGACTCAAGACGATAGTTACCGGATAAGGCGCAGCGGTCGGGC
TGAACGGGGGGTTCGTGCACACAGCCCAGCTTGGAGCGAACGACCTACACCGAACTGAGA
TACCTACAGCGTGAGCTATGAGAAAGCGCCACGCTTCCCGAAGGGAGAAAGGCGGACAGG
TATCCGGTAAGCGGCAGGGTCGGAACAGGAGAGCGCACGAGGGAGCTTCCAGGGGGAAAC
GCCTGGTATCTTTATAGTCCTGTCGGGTTTCGCCACCTCTGACTTGAGCGTCGATTTTTG
TGATGCTCGTCAGGGGGGCGGAGCCTATGGAAAAACGCCAGCAACGCGGCCTTTTTACGG
TTCCTGGCCTTTTGCTGGCCTTTTGCTCACATGTTCTTTCCTGCGTTATCCCCTGATTCT
GTGGATAACCGTATTACCGCCTTTGAGTGAGCTGATACCGCTCGCCGCAGCCGAACGACC
GAGCGCAGCGAGTCAGTGAGCGAGGAAGCGGAAGAGCGCCCAATACGCAAACCGCCTCTC
CCCGCGCGTTGGCCGATTCATTAATGCAGCTGGCACGACAGGTTTCCCGACTGGAAAGCG
GGCAGTGAGCGCAACGCAATTAATGTGAGTTAGCTCACTCATTAGGCACCCCAGGCTTTA
CACTTTATGCTTCCGGCTCGTATGTTGTGTGGAATTGTGA
Restriction sites to linearize plasmid
Partial or complete disruption of the geneComplete
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 present in oocyst-derived sporozoites and not in salivary gland sporozoites. It contains a signal peptide, a transmembrane region and a PEXEL motif.
The gentic modification results in the disruption of the following gene models PB000015.03.0, PB301066.00.0 and PB301121.00.0. See for more information about the disruption Lasonder, E. et al., 2008, PloS Pathogens 10, e10000195 (Supporting Information: S3).

Independent unsuccessful attempts to disrupt this gene has been reported (see RMgm190.
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 1AAAGGTACCTTCACTAGCGTAATTCTG
Additional information primer 1L2656 (Asp718); 5'
Sequence Primer 2AAAAAGCTTGCTATGATGATTTTCG
Additional information primer 2L2657 (HindIII); 5'
Sequence Primer 3AAAGAATTCGTGAATTATATTAACAGC
Additional information primer 3L2658 (EcoRI); 3'
Sequence Primer 4AAAGCGGCCGCATTAACATATGTGTTACC
Additional information primer 4L2659 (NotI); 3'
Sequence Primer 5
Additional information primer 5
Sequence Primer 6
Additional information primer 6