Summary

RMgm-250
Malaria parasiteP. berghei
Genotype
Genetic modification not successful
DisruptedGene model (rodent): PBANKA_1402600; Gene model (P.falciparum): PF3D7_1304100; Gene product: DNA ligase I (PfLig1)
PhenotypeNo phenotype has been described
Last modified: 4 May 2009, 21:34
  *RMgm-250
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 4
Reference (PubMed-PMID number) Not published (yet)
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).
Attempts to generate the mutant parasite were performed by
Name PI/ResearcherG.R. Mair, S.M. Khan, C.J. Janse
Name Group/DepartmentLeiden Malaria Research Group
Name InstituteLeiden University Medical Center
CityLeiden
CountryThe Netherlands

  Disrupted: Mutant parasite with a disrupted gene
Details of the target gene
Gene Model of Rodent Parasite PBANKA_1402600
Gene Model P. falciparum ortholog PF3D7_1304100
Gene productDNA ligase I
Gene product: Alternative namePfLig1
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
Click to view information
Click to hide information
Plasmid/construct sequence
Click to view information
Click to hide information
AATTCAAATTTTTTAATGCCAAGGAGCAAATTATTTTATTATACATTTTATTTTTTTTTA
ATGCGTGTTTCTCATTTTCTATTAATTTTGCATTTTTATTTACAATTAATATTCTTATTA
TCCTTATTTTATTTTGTGTCGTCTTTCTCATTTATTTCGCCATTTTTTATATATTTTTTA
ATCCTATGTAATTTTTACACATTTCAGCATCTGTGAAAATGAGATGTGTATAGTAAAAAA
AATATATACAAAGATCCTATTTGAACTAATTATTTAATAATTTACACAAAATTTTAAAAT
ATAAATTAACAATTTATTCATATATGTTTTTTTCCAATTTTTTAATTATTTTTTACTATA
AAATGTAGAAATTTATTAATTTTAATCAACATAATATGATTTAAATAAGCCTAAGGGGTA
AATGCATATAATGTAAATACTTCTTATATGTATATTTATGATTTATATACCTTGAGATGT
ATTTATTTTTTAATACATATATATATATATCAATAAATATAAATAAAATAAGCATATACA
TATTGTTTATATTTATACTTATTTAAAATTGGCTTATTTTTAATTATTATATGAATTATG
GAAAAGCCACTTAAGCAAGTAACCTTAGAGTGAGTTAAATAAAAAAAATTTATGATTACA
TAAATAATGGATGTTATTGTAAATAATATAGGAAATATTAATGATATTTAATACTTATTA
TCTTTTTACGTAGCAATTTTTATTGCAAAATCATATTATTTTGGTGCTGATCGAATATGC
AGGAAATGCAGCCATATGCATATTTGCATGCGTAGTTTATGTTATTGATTGAAATATTTT
TTGGTGTAATATTTTTTCAAATTGGGGGGGAAATGAATGTATATAATGTTGGCATAAGTA
TTTTTGAATAGTTATTAAAAAATATGATGCATAAGAATAATGTGCATACATGTAACAATA
CGAATACGCATGGATACATTAATAATATATGTCAATTTAATATGCAATATATTTTTTTTT
GTATTTTGCTAATGAAATGCTCTTCATTGTAGCATTAACTAGCGGTTAATATTGAATTAT
TTGTCTTGTGTATTTGTATTACTTAAGCTTGCATGCCTGCAGGTCAACAATAAATAATAA
ATAAATATTGTGGAAATAAAATAACATATAATTATTTTTAATACATTGATTTCCCTTTTA
TTTTTTTAAATTTCATTGATATAAAAATATATAATAATAACATATATGATTTCAAATTAA
TCTTTTCAAAAATGGTGTTTATTTTTGTATGTTTGTGTATGAATTAATCACATAACACAT
CTATTAAATTGAGTTGGTAATATAGACACAAATAAATATATATATTTTTATAGCTTAAAA
GTGTGTTATGAATATTTTAAGCATATTTTCTTTTTCTTTGGATTGTGTAAAATGAACTCA
TATAATGCGTTTTTTTGTTTTTGTTATTTTGTCATTTTGTTATTTTGCTATTTTATGGAT
TAATTTTTGTTTATAAAATGGGAAATAATTTTAACATATTTAAATAAATGGAGAAAAAAT
ATAAAATAATTATAAAAAAAAGTTAATACACATTTTTTCCTGTTATAGACCTTATATTTA
TTTATCCATATATATATATATATATATATATATATATATATACATACCAAGTGAATTAAG
AGGAAAGCTAATTTATTATTCAGAATAATATATGAACTATATATAATTTTTATTATTTTG
GTGTATATTAATCTGTCTATATGCATACATGCAATAATTTATCGACTTATATATCAAATA
ACATAAAATAGAAGTGTTTTAAATTATGGATATATGCTCAATATTCATTTTTTTTAATAA
GTTAGCTATATTTAAATTATACATTTTATATATGGTCTCTTTTTTTTTTAAATATTATTT
AAGTGATCATGAAAATATAAATAATTTTTTTTTATTTAATATCCTTTTGCTTGCATGTGG
TAAATGGAAATTTGGATGTGTTTTGAARGTTCGGATATAGTTGTATGGACATATAATATA
TTTTGTGAAAAATTGGTTTTATGTTTATACTTATGCCAATACTTTTTGAGTAAAACAAAG
CAAGTGCTTATAAATAATTAAAGCCAATTTTATAATATATATTTTTTTATTTAATTTGAA
TTTAGTAGTATAATTTTTTATGGTAAGTGCTCAAAGAGAGTTGCTTATAAAGTATGGTTT
GTTTCTTTTTCGCCATTTTGAATTACACATTAAAAATATATAGATACATATATTATAATA
TGAAATCATTAATAATTTAGGGAAATTCTACAAATTTAAAAACGAATAAAATAATTGTTT
TTCATCATGCCATAACACAATATTGATATATACATGTACAAACATTTTTTTTATTTGGAA
AATATAAATTATATAAAAAAAAATGTATAGTATACAAAATGAGCATATTCACACGGGGTG
GACGTTCATTTTTTCATTTTTCCCCTGTTTTTTATGAGTATATGATAAAATTTTATGAAC
ATTTACACAAAATGAAAATGGATATATAGGAAAAATGGAGCGGTATTTCATTTATCTTTG
ATTGTCATTTGGATATTATATTACCYTGGGTAGGCAATTAAAAATGTTAAATAACAATTT
AAGGAAATTATATTTTATATATTAAAATTAACACTGTATTATATGATTCGCTTATAAAAG
CCACTCTTTCCCCATGCAAAGCTGTTTAATATCAATTTTAACAAATTACACACATGTTAA
TATATTTATATATATAATTTATATATTTATAATTTATATATTTATATTTTTATTATTTAT
ATATTTATTATTTATTGTGTGTGTCAATTCGGGTAGGATATACCTCTTTTTTATTGTTTA
AAGCGATTTGTATTCTAAAATATAAAGRATTTGAAAAAGAGAAAGATAGAATATGATCCC
ATCATATATAGCCCTATAATTTTTATTTAGCAGCGAATTAATTTTTCTATTAAGTTTATG
TGTAATTAAAATAACGGAATATATATAATACAATAAAAAAGTGCATAAATTAAAATTTTT
TCAATTAAATTTTTTTTTTTAAGGGGTTATATAATATTAAATATATAAAATACGATTATA
TATTTTTGCTACAATTTTTTATATTAAGATATAAATAGTAAATAAATGGTATTATATGGC
ATGTAATATATAAATTTTTTCCAATTTTTATTTTATATACACTTTTCCTTTTTTTGTCAT
AAAACTTAAACAATTTACACATTCATTTTAAAAATTGACTATTTGTTTCAACATTTTTTG
AGTTTCCGTTTTATAATAGTATTTTCATTTGTATATTGCTTATATATATAAATACACACC
TAAATGTTACAAAGGATCAATGCATAAACCGGTGTGTCTGGTCGTCGCGATGACCCCCAA
GAGGGGCATCGGCATCAACAACGGCCTCCCGTGGCCCCACTTGACCACAGATTTCAAACA
CTTTTCTCGTGTGACAAAAACGACGCCCGAAGAAGCCAGTCGCCTGAACGGGTGGCTTCC
CAGGAAATTTGCAAAGACGGGCGACTCTGGACTTCCCTCTCCATCAGTCGGCAAGAGATT
CAACGCCGTTGTCATGGGACGGAAAACCTGGGAAAGCATGCCTCGAAAGTTTAGACCCCT
CGTGGACAGATTGAACATCGTCGTTTCCTCTTCCCTCAAAGAAGAAGACATTGCGGCGGA
GAAGCCTCAAGCTGAAGGCCAGCAGCGCGTCCGAGTCTGTGCTTCACTCCCAGCAGCTCT
CAGCCTTCTGGAGGAAGAGTACAAGGATTCTGTCGACCAGATTTTTGTCGTGGGAGGAGC
GGGACTGTACGAGGCAGCGCTGTCTCTGGGCGTTGCCTCTCACCTGTACATCACGCGTGT
AGCCCGCGAGTTTCCGTGCGACGTTTTCTTCCCTGCGTTCCCCGGAGATGACATTCTTTC
AAACAAATCAACTGCTGCGCAGGCTGCAGCTCCTGCCGAGTCTGTGTTCGTTCCCTTTTG
TCCGGAGCTCGGAAGAGAGAAGGACAATGAAGCGACGTATCGACCCATCTTCATTTCCAA
GACCTTCTCAGACAACGGGGTTCCCTACGACTTTGTGGTTCTCGAGAAGAGAAGGAAGAC
TGACGACGCAGCCACTGCGGAACCGAGCAACGCAATGAGCTCCTTGACGTCCACGAGGGA
GACAACTCCCGTGCACGGGTTGCAGGCTCCTTCTTCGGCCGCAGCCATTGCCCCGGTGTT
GGCGTGGATGGACGAAGAAGACCGGAAAAAACGCGAGCAAAAGGAACTGATTCGGGCCGT
TCCGCATGTTCACTTTAGAGGCCATGAAGAGTTCCAGTACCTTGATCTCATTGCCGACAT
TATTAACAATGGAAGGACAATGGATGACCGAACGGGCGTTGGTGTCATCTCCAAATTCGG
CTGCACTATGCGCTACTCGCTGGATCAGGCCTTTCCACTTCTCACCACAAAGCGTGTGTT
CTGGAAAGGGGTCCTCGAAGAGTTGCTGTGGTTCATTCGCGGCGACACGAACGCAAACCA
TCTTTCTGAGAAGGGCGTGAAGATCTGGGACAAGAATGTGACACGCGAGTTCCTCGATTC
GCGCAATCTCCCCCACCGAGAGGTCGGAGACATCGGCCCGGGCTACGGCTTCCAGTGGAG
ACACTTCGGCGCGGCATACAAAGACATGCACACAGACTACACAGGGCAGGGCGTCGACCA
GCTGAAGAATGTGATCCAGATGCTGAGAACGAATCCAACAGATCGTCGCATGCTCATGAC
TGCCTGGAATCCTGCAGCGCTGGACGAAATGGCGCTGCCGCCTTGTCACTTGTTGTGCCA
GTTCTACGTGAACGACCAGAAGGAGCTGTCGTGCATCATGTATCAGCGGTCGTGCGATGT
CGGCCTCGGCGTCCCCTTCAACATCGCTTCCTATTCGCTTTTGACGCTCATGGTTGCACA
CGTCTGCAACCTAAAACCTAAGGAGTTCATTCACTTCATGGGGAACACGCATGTCTACAC
GAACCATGTCGAGGCTTTAAAAGAGCAGCTGCGGAGAGAACCGAGACCGTTCCCCATTGT
GAACATCCTCAACAAGGAACGCATCAAGGAAATCGACGATTTCACCGCCGAGGATTTTGA
GGTCGTGGGCTACGTCCCGCACGGACGAATCCAGATGGAGATGGCTGTCTAGCGGAAATA
CAGAAGCTAGCTTTGATCCCGTTTTTCTTACTTATATATTTATACCAATTGATTGTATTT
ATAACTGTAAAAATGTGTATGTTGTGTGCATATTTTTTTTTGTGCATGCACATGCATGTA
AATAGCTAAAATTATGAACATTTTATTTTTTGTTCAGAAAAAAAAAACTTTACACACATA
AAATGGCTAGTATGAATAGCCATATTTTATATAAATTAAATCCTATGAATTTATGACCAT
ATTAAAAATTTAGATATTTATGGAACATAATATGTTTGAAACAATAAGACAAAATTATTA
TTATTATTATTATTTTTACTGTTATAATTATGTTGTCTCTTCAATGATTCATAAATAGTT
GGACTTGATTTTTAAAATGTTTATAATATGATTAGCATAGTTAAATAAAAAAAGTTGAAA
AATTAAAAAAAAACATATAAACACAAATGATGTTTTTTCCTTCAATTTCGATATCGATTG
GGTTAAGATTCCCAAGGTTTCTTAGATTAAGGGATGATAAGAATCCAGAACAAGCAACAA
CAGCTCAGCAGATTGTAGAATTTTACGAAGCTCAATTTGCCTCGAAGAAAAATAAAAATA
ACAATTATAACGACGAAAGTGGAAGCGAATAGTTTTATAACTCTGGGGTGAAGGCGACAT
ATATGTATGCGTATTTTTTATACACTTACATATATGCATGTTTCTATATATTAATATAAG
CGCATTCACACCCGAACTGAATCAATTCTTTTATTCGAAAACAAAATCATAATATATATA
TGTGTACATTAGTATGGAAATGTAACATAATATTTTTTTAATATAGTTTCAAATTTATAC
CGATAATATCGTCTATATAGACAATGTTTATTTTTATTTAATAGCTCTTTTTGCAACTTT
AAAAAATATTTCTTATAAAAAAATAAACTCTGCATTTTTGCATTTTATTTGTTTTTTTTT
GTTTTAAAATACCCATAAAGTTATATGTATATATTGAAACAGTTATGTACAACTATATCC
ACTTTAAATAAACTTTTATGAATAAATAAATCACTAATTCCAAATAGGTGTTAGGCTACG
ATAATTGAATATAAGAAATGTGATACATATATATATGGAATATATAAGGATTTATTTTGG
ATCCACTAGTTCTAGAGCGGCCGCCACCGCGGTGGAGCTCCAATTCGCCCTATAGTGAGT
CGTATTACGCGCGCTCACTGGCCGTCGTTTTACAACGTCGTGACTGGGAAAACCCTGGCG
TTACCCAACTTAATCGCCTTGCAGCACATCCCCCTTTCGCCAGCTGGCGTAATAGCGAAG
AGGCCCGCACCGATCGCCCTTCCCAACAGTTGCGCAGCCTGAATGGCGAATGGGACGCGC
CCTGTAGCGGCGCATTAAGCGCGGCGGGTGTGGTGGTTACGCGCAGCGTGACCGCTACAC
TTGCCAGCGCCCTAGCGCCCGCTCCTTTCGCTTTCTTCCCTTCCTTTCTCGCCACGTTCG
CCGGCTTTCCCCGTCAAGCTCTAAATCGGGGGCTCCCTTTAGGGTTCCGATTTAGTGCTT
TACGGCACCTCGACCCCAAAAAACTTGATTAGGGTGATGGTTCACGTAGTGGGCCATCGC
CCTGATAGACGGTTTTTCGCCCTTTGACGTTGGAGTCCACGTTCTTTAATAGTGGACTCT
TGTTCCAAACTGGAACAACACTCAACCCTATCTCGGTCTATTCTTTTGATTTATAAGGGA
TTTTGCCGATTTCGGCCTATTGGTTAAAAAATGAGCTGATTTAACAAAAATTTAACGCGA
ATTTTAACAAAATATTAACGCTTACAATTTAGGTGGCACTTTTCGGGGAAATGTGCGCGG
AACCCCTATTTGTTTATTTTTCTAAATACATTCAAATATGTATCCGCTCATGAGACAATA
ACCCTGATAAATGCTTCAATAATATTGAAAAAGGAAGAGTATGAGTATTCAACATTTCCG
TGTCGCCCTTATTCCCTTTTTTGCGGCATTTTGCCTTCCTGTTTTTGCTCACCCAGAAAC
GCTGGTGAAAGTAAAAGATGCTGAAGATCAGTTGGGTGCACGAGTGGGTTACATCGAACT
GGATCTCAACAGCGGTAAGATCCTTGAGAGTTTTCGCCCCGAAGAACGTTTTCCAATGAT
GAGCACTTTTAAAGTTCTGCTATGTGGCGCGGTATTATCCCGTATTGACGCCGGGCAAGA
GCAACTCGGTCGCCGCATACACTATTCTCAGAATGACTTGGTTGAGTACTCACCAGTCAC
AGAAAAGCATCTTACGGATGGCATGACAGTAAGAGAATTATGCAGTGCTGCCATAACCAT
GAGTGATAACACTGCGGCCAACTTACTTCTGACAACGATCGGAGGACCGAAGGAGCTAAC
CGCTTTTTTGCACAACATGGGGGATCATGTAACTCGCCTTGATCGTTGGGAACCGGAGCT
GAATGAAGCCATACCAAACGACGAGCGTGACACCACGATGCCTGTAGCAATGGCAACAAC
GTTGCGCAAACTATTAACTGGCGAACTACTTACTCTAGCTTCCCGGCAACAATTAATAGA
CTGGATGGAGGCGGATAAAGTTGCAGGACCACTTCTGCGCTCGGCCCTTCCGGCTGGCTG
GTTTATTGCTGATAAATCTGGAGCCGGTGAGCGTGGGTCTCGCGGTATCATTGCAGCACT
GGGGCCAGATGGTAAGCCCTCCCGTATCGTAGTTATCTACACGACGGGGAGTCAGGCAAC
TATGGATGAACGAAATAGACAGATCGCTGAGATAGGTGCCTCACTGATTAAGCATTGGTA
ACTGTCAGACCAAGTTTACTCATATATACTTTAGATTGATTTAAAACTTCATTTTTAATT
TAAAAGGATCTAGGTGAAGATCCTTTTTGATAATCTCATGACCAAAATCCCTTAACGTGA
GTTTTCGTTCCACTGAGCGTCAGACCCCGTAGAAAAGATCAAAGGATCTTCTTGAGATCC
TTTTTTTCTGCGCGTAATCTGCTGCTTGCAAACAAAAAAACCACCGCTACCAGCGGTGGT
TTGTTTGCCGGATCAAGAGCTACCAACTCTTTTTCCGAAGGTAACTGGCTTCAGCAGAGC
GCAGATACCAAATACTGTCCTTCTAGTGTAGCCGTAGTTAGGCCACCACTTCAAGAACTC
TGTAGCACCGCCTACATACCTCGCTCTGCTAATCCTGTTACCAGTGGCTGCTGCCAGTGG
CGATAAGTCGTGTCTTACCGGGTTGGACTCAAGACGATAGTTACCGGATAAGGCGCAGCG
GTCGGGCTGAACGGGGGGTTCGTGCACACAGCCCAGCTTGGAGCGAACGACCTACACCGA
ACTGAGATACCTACAGCGTGAGCTATGAGAAAGCGCCACGCTTCCCGAAGGGAGAAAGGC
GGACAGGTATCCGGTAAGCGGCAGGGTCGGAACAGGAGAGCGCACGAGGGAGCTTCCAGG
GGGAAACGCCTGGTATCTTTATAGTCCTGTCGGGTTTCGCCACCTCTGACTTGAGCGTCG
ATTTTTGTGATGCTCGTCAGGGGGGCGGAGCCTATGGAAAAACGCCAGCAACGCGGCCTT
TTTACGGTTCCTGGCCTTTTGCTGGCCTTTTGCTCACATGTTCTTTCCTGCGTTATCCCC
TGATTCTGTGGATAACCGTATTACCGCCTTTGAGTGAGCTGATACCGCTCGCCGCAGCCG
AACGACCGAGCGCAGCGAGTCAGTGAGCGAGGAAGCGGAAGAGCGCCCAATACGCAAACC
GCCTCTCCCCGCGCGTTGGCCGATTCATTAATGCAGCTGGCACGACAGGTTTCCCGACTG
GAAAGCGGGCAGTGAGCGCAACGCAATTAATGTGAGTTAGCTCACTCATTAGGCACCCCA
GGCTTTACACTTTATGCTTCCGGCTCGTATGTTGTGTGGAATTGTGAGCGGATAACAATT
TCACACAGGAAACAGCTATGACCATGATTACGCCAAGCTCGAAATTAACCCTCACTAAAG
GGAACAAAAGCTGGTACCGGGCCCCCCCTCGAGGTCGACGGTATCGATG
Restriction sites to linearize plasmid
Partial or complete disruption of the geneComplete
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 modificationDisruption experiments were performed in the Leiden Malaria Research Group (exp. 548, 585, 609, 629; pl1026).
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 1aaagaattcAAATTTTTTAATGCCAAGGAGC
Additional information primer 1L2226 (EcoRI); 5'
Sequence Primer 2aaaaagcttAAGTAATACAAATACACAAGAC
Additional information primer 2L2227 (HindIII); 5'
Sequence Primer 3aaagatatcGATTGGGTTAAGATTCCCAAGG
Additional information primer 3L2228 (EcoRV); 3'
Sequence Primer 4aaaggatccAAAATAAATCCTTATATATTCC
Additional information primer 4L2229 (BamHI); 3'
Sequence Primer 5
Additional information primer 5
Sequence Primer 6
Additional information primer 6