Summary

RMgm-261
Malaria parasiteP. berghei
Genotype
DisruptedGene model (rodent): PBANKA_1231300; Gene model (P.falciparum): PF3D7_0516500; Gene product: major facilitator superfamily domain-containing protein, putative
PhenotypeNo phenotype has been described
Last modified: 6 May 2009, 17:12
  *RMgm-261
Successful modificationThe parasite was generated by the genetic modification
The mutant contains the following genetic modification(s) Gene disruption
Reference (PubMed-PMID number) Not published (yet)
MR4 number
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).
The mutant parasite was generated by
Name PI/ResearcherJ. Kroeze, C.J. Janse, A.P. Waters
Name Group/DepartmentLeiden Malaria Research Group
Name InstituteLeiden University Medical Center
CityLeiden
CountryThe Netherlands
Name of the mutant parasite
RMgm numberRMgm-261
Principal name648cl1,cl3 (680 uncloned)
Alternative name
Standardized name
Is the mutant parasite cloned after genetic modificationYes
Phenotype
Asexual blood stageNot different from wild type
Gametocyte/GameteNot different from wild type
Fertilization and ookineteNot different from wild type
OocystNot different from wild type
SporozoiteNot different from wild type
Liver stageNot different from wild type
Additional remarks phenotype

Mutant/mutation
The mutant lacks expression of PB000201.02.0.

Protein (function)

Phenotype
The phenotype has not been analysed in detail. Blood stage development and gametocyte and ookinete production were comparable to those of wild type.  Oocyst and sporozoite production and prepatent period after infection of mice by mosquito bite was comparable to those of wild type parasites. See also 'Additional remarks partial/complete disruption'.

The mutant can be obtained from the Leiden Malaria Research Group.

 


  Disrupted: Mutant parasite with a disrupted gene
Details of the target gene
Gene Model of Rodent Parasite PBANKA_1231300
Gene Model P. falciparum ortholog PF3D7_0516500
Gene productmajor facilitator superfamily domain-containing protein, putative
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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CCCCCTCGAGGTCGACGGTATCGATAAGCTTGCATGCCTGCAGGTCAACAATAAATAATA
AATAAATATTGTGGAAATAAAATAACATATAATTATTTTTAATACATTGATTTCCCTTTT
ATTTTTTTAAATTTCATTGATATAAAAATATATAATAATAACATATATGATTTCAAATTA
ATCTTTTCAAAAATGGTGTTTATTTTTGTATGTTTGTGTATGAATTAATCACATAACACA
TCTATTAAATTGAGTTGGTAATATAGACACAAATAAATATATATATTTTTATAGCTTAAA
AGTGTGTTATGAATATTTTAAGCATATTTTCTTTTTCTTTGGATTGTGTAAAATGAACTC
ATATAATGCGTTTTTTTGTTTTTGTTATTTTGTCATTTTGTTATTTTGCTATTTTATGGA
TTAATTTTTGTTTATAAAATGGGAAATAATTTTAACATATTTAAATAAATGGAGAAAAAA
TATAAAATAATTATAAAAAAAAGTTAATACACATTTTTTCCTGTTATAGACCTTATATTT
ATTTATCCATATATATATATATATATATATATATATATATATACATACCAAGTGAATTAA
GAGGAAAGCTAATTTATTATTCAGAATAATATATGAACTATATATAATTTTTATTATTTT
GGTGTATATTAATCTGTCTATATGCATACATGCAATAATTTATCGACTTATATATCAAAT
AACATAAAATAGAAGTGTTTTAAATTATGGATATATGCTCAATATTCATTTTTTTTAATA
AGTTAGCTATATTTAAATTATACATTTTATATATGGTCTCTTTTTTTTTTAAATATTATT
TAAGTGATCATGAAAATATAAATAATTTTTTTTTATTTAATATCCTTTTGCTTGCATGTG
GTAAATGGAAATTTGGATGTGTTTTGAARGTTCGGATATAGTTGTATGGACATATAATAT
ATTTTGTGAAAAATTGGTTTTATGTTTATACTTATGCCAATACTTTTTGAGTAAAACAAA
GCAAGTGCTTATAAATAATTAAAGCCAATTTTATAATATATATTTTTTTATTTAATTTGA
ATTTAGTAGTATAATTTTTTATGGTAAGTGCTCAAAGAGAGTTGCTTATAAAGTATGGTT
TGTTTCTTTTTCGCCATTTTGAATTACACATTAAAAATATATAGATACATATATTATAAT
ATGAAATCATTAATAATTTAGGGAAATTCTACAAATTTAAAAACGAATAAAATAATTGTT
TTTCATCATGCCATAACACAATATTGATATATACATGTACAAACATTTTTTTTATTTGGA
AAATATAAATTATATAAAAAAAAATGTATAGTATACAAAATGAGCATATTCACACGGGGT
GGACGTTCATTTTTTCATTTTTCCCCTGTTTTTTATGAGTATATGATAAAATTTTATGAA
CATTTACACAAAATGAAAATGGATATATAGGAAAAATGGAGCGGTATTTCATTTATCTTT
GATTGTCATTTGGATATTATATTACCYTGGGTAGGCAATTAAAAATGTTAAATAACAATT
TAAGGAAATTATATTTTATATATTAAAATTAACACTGTATTATATGATTCGCTTATAAAA
GCCACTCTTTCCCCATGCAAAGCTGTTTAATATCAATTTTAACAAATTACACACATGTTA
ATATATTTATATATATAATTTATATATTTATAATTTATATATTTATATTTTTATTATTTA
TATATTTATTATTTATTGTGTGTGTCAATTCGGGTAGGATATACCTCTTTTTTATTGTTT
AAAGCGATTTGTATTCTAAAATATAAAGRATTTGAAAAAGAGAAAGATAGAATATGATCC
CATCATATATAGCCCTATAATTTTTATTTAGCAGCGAATTAATTTTTCTATTAAGTTTAT
GTGTAATTAAAATAACGGAATATATATAATACAATAAAAAAGTGCATAAATTAAAATTTT
TTCAATTAAATTTTTTTTTTTAAGGGGTTATATAATATTAAATATATAAAATACGATTAT
ATATTTTTGCTACAATTTTTTATATTAAGATATAAATAGTAAATAAATGGTATTATATGG
CATGTAATATATAAATTTTTTCCAATTTTTATTTTATATACACTTTTCCTTTTTTTGTCA
TAAAACTTAAACAATTTACACATTCATTTTAAAAATTGACTATTTGTTTCAACATTTTTT
GAGTTTCCGTTTTATAATAGTATTTTCATTTGTATATTGCTTATATATATAAATACACAC
CTAAATGTTACAAAGGATCAATGCATAAACCGGTGTGTCTGGTCGTCGCGATGACCCCCA
AGAGGGGCATCGGCATCAACAACGGCCTCCCGTGGCCCCACTTGACCACAGATTTCAAAC
ACTTTTCTCGTGTGACAAAAACGACGCCCGAAGAAGCCAGTCGCCTGAACGGGTGGCTTC
CCAGGAAATTTGCAAAGACGGGCGACTCTGGACTTCCCTCTCCATCAGTCGGCAAGAGAT
TCAACGCCGTTGTCATGGGACGGAAAACCTGGGAAAGCATGCCTCGAAAGTTTAGACCCC
TCGTGGACAGATTGAACATCGTCGTTTCCTCTTCCCTCAAAGAAGAAGACATTGCGGCGG
AGAAGCCTCAAGCTGAAGGCCAGCAGCGCGTCCGAGTCTGTGCTTCACTCCCAGCAGCTC
TCAGCCTTCTGGAGGAAGAGTACAAGGATTCTGTCGACCAGATTTTTGTCGTGGGAGGAG
CGGGACTGTACGAGGCAGCGCTGTCTCTGGGCGTTGCCTCTCACCTGTACATCACGCGTG
TAGCCCGCGAGTTTCCGTGCGACGTTTTCTTCCCTGCGTTCCCCGGAGATGACATTCTTT
CAAACAAATCAACTGCTGCGCAGGCTGCAGCTCCTGCCGAGTCTGTGTTCGTTCCCTTTT
GTCCGGAGCTCGGAAGAGAGAAGGACAATGAAGCGACGTATCGACCCATCTTCATTTCCA
AGACCTTCTCAGACAACGGGGTTCCCTACGACTTTGTGGTTCTCGAGAAGAGAAGGAAGA
CTGACGACGCAGCCACTGCGGAACCGAGCAACGCAATGAGCTCCTTGACGTCCACGAGGG
AGACAACTCCCGTGCACGGGTTGCAGGCTCCTTCTTCGGCCGCAGCCATTGCCCCGGTGT
TGGCGTGGATGGACGAAGAAGACCGGAAAAAACGCGAGCAAAAGGAACTGATTCGGGCCG
TTCCGCATGTTCACTTTAGAGGCCATGAAGAGTTCCAGTACCTTGATCTCATTGCCGACA
TTATTAACAATGGAAGGACAATGGATGACCGAACGGGCGTTGGTGTCATCTCCAAATTCG
GCTGCACTATGCGCTACTCGCTGGATCAGGCCTTTCCACTTCTCACCACAAAGCGTGTGT
TCTGGAAAGGGGTCCTCGAAGAGTTGCTGTGGTTCATTCGCGGCGACACGAACGCAAACC
ATCTTTCTGAGAAGGGCGTGAAGATCTGGGACAAGAATGTGACACGCGAGTTCCTCGATT
CGCGCAATCTCCCCCACCGAGAGGTCGGAGACATCGGCCCGGGCTACGGCTTCCAGTGGA
GACACTTCGGCGCGGCATACAAAGACATGCACACAGACTACACAGGGCAGGGCGTCGACC
AGCTGAAGAATGTGATCCAGATGCTGAGAACGAATCCAACAGATCGTCGCATGCTCATGA
CTGCCTGGAATCCTGCAGCGCTGGACGAAATGGCGCTGCCGCCTTGTCACTTGTTGTGCC
AGTTCTACGTGAACGACCAGAAGGAGCTGTCGTGCATCATGTATCAGCGGTCGTGCGATG
TCGGCCTCGGCGTCCCCTTCAACATCGCTTCCTATTCGCTTTTGACGCTCATGGTTGCAC
ACGTCTGCAACCTAAAACCTAAGGAGTTCATTCACTTCATGGGGAACACGCATGTCTACA
CGAACCATGTCGAGGCTTTAAAAGAGCAGCTGCGGAGAGAACCGAGACCGTTCCCCATTG
TGAACATCCTCAACAAGGAACGCATCAAGGAAATCGACGATTTCACCGCCGAGGATTTTG
AGGTCGTGGGCTACGTCCCGCACGGACGAATCCAGATGGAGATGGCTGTCTAGCGGAAAT
ACAGAAGCTAGCTTTGATCCCGTTTTTCTTACTTATATATTTATACCAATTGATTGTATT
TATAACTGTAAAAATGTGTATGTTGTGTGCATATTTTTTTTTGTGCATGCACATGCATGT
AAATAGCTAAAATTATGAACATTTTATTTTTTGTTCAGAAAAAAAAAACTTTACACACAT
AAAATGGCTAGTATGAATAGCCATATTTTATATAAATTAAATCCTATGAATTTATGACCA
TATTAAAAATTTAGATATTTATGGAACATAATATGTTTGAAACAATAAGACAAAATTATT
ATTATTATTATTATTTTTACTGTTATAATTATGTTGTCTCTTCAATGATTCATAAATAGT
TGGACTTGATTTTTAAAATGTTTATAATATGATTAGCATAGTTAAATAAAAAAAGTTGAA
AAATTAAAAAAAAACATATAAACACAAATGATGTTTTTTCCTTCAATTTCGATATCGCTG
CTATGTGCTTTCATTTGTTTTGTCATCACAATATACATTTTTGTATTAGATCAATCGACG
CTGAAAATTTTTGAAAACGATGCGGATAGTTTAGAGACCATGTTTTCTAGTATAAAACTA
TCATTGTAGTATATTTTTGTCAATTGTTTTTTTCCGTTTTTTAATCGCAAATTTTGTATC
AATATGCGCCATAAAATTTATTTGAAAAAATAAATAATTTTGCTGTACTATTTCTTTTGC
ATTTTATTGTCTTTTTTTTTAATATTTTTTTACTTAATCGTAATCCGTGATATAAGGAAT
ACACCTTATTGGGTTTGAAATAAGTTGATCTTTTTCACAAAAAAATTTTAGCAAATGAAA
AGATGTGCATTTAGCACGAATTTTTTGGTGTAAATAAAATAGGTGGATAAGGATTGATAA
ATAAATAGTATTAACAAAGCCAACGAACATACAAACATCTTAAACAAAATTAACAAATAA
AAGAATAAAAAATGTAATAAAAAATAAAAAAAATGATTTTCAAGTTAGATATTAAGACAA
TTAAAAAATCAGTATGCACATAAGCAGGATTGTGCAAAAGTGAATAGTGGTTTTAAAAAA
TTAAAATTCGCACACTACTCAATTATCGCGATGGCGCTGAATCAATTGTTCGTAAATAAT
TGCGGATCCACTAGTTCTAGAGCGGCCGCCACCGCGGTGGAGCTCCAATTCGCCCTATAG
TGAGTCGTATTACGCGCGCTCACTGGCCGTCGTTTTACAACGTCGTGACTGGGAAAACCC
TGGCGTTACCCAACTTAATCGCCTTGCAGCACATCCCCCTTTCGCCAGCTGGCGTAATAG
CGAAGAGGCCCGCACCGATCGCCCTTCCCAACAGTTGCGCAGCCTGAATGGCGAATGGGA
CGCGCCCTGTAGCGGCGCATTAAGCGCGGCGGGTGTGGTGGTTACGCGCAGCGTGACCGC
TACACTTGCCAGCGCCCTAGCGCCCGCTCCTTTCGCTTTCTTCCCTTCCTTTCTCGCCAC
GTTCGCCGGCTTTCCCCGTCAAGCTCTAAATCGGGGGCTCCCTTTAGGGTTCCGATTTAG
TGCTTTACGGCACCTCGACCCCAAAAAACTTGATTAGGGTGATGGTTCACGTAGTGGGCC
ATCGCCCTGATAGACGGTTTTTCGCCCTTTGACGTTGGAGTCCACGTTCTTTAATAGTGG
ACTCTTGTTCCAAACTGGAACAACACTCAACCCTATCTCGGTCTATTCTTTTGATTTATA
AGGGATTTTGCCGATTTCGGCCTATTGGTTAAAAAATGAGCTGATTTAACAAAAATTTAA
CGCGAATTTTAACAAAATATTAACGCTTACAATTTAGGTGGCACTTTTCGGGGAAATGTG
CGCGGAACCCCTATTTGTTTATTTTTCTAAATACATTCAAATATGTATCCGCTCATGAGA
CAATAACCCTGATAAATGCTTCAATAATATTGAAAAAGGAAGAGTATGAGTATTCAACAT
TTCCGTGTCGCCCTTATTCCCTTTTTTGCGGCATTTTGCCTTCCTGTTTTTGCTCACCCA
GAAACGCTGGTGAAAGTAAAAGATGCTGAAGATCAGTTGGGTGCACGAGTGGGTTACATC
GAACTGGATCTCAACAGCGGTAAGATCCTTGAGAGTTTTCGCCCCGAAGAACGTTTTCCA
ATGATGAGCACTTTTAAAGTTCTGCTATGTGGCGCGGTATTATCCCGTATTGACGCCGGG
CAAGAGCAACTCGGTCGCCGCATACACTATTCTCAGAATGACTTGGTTGAGTACTCACCA
GTCACAGAAAAGCATCTTACGGATGGCATGACAGTAAGAGAATTATGCAGTGCTGCCATA
ACCATGAGTGATAACACTGCGGCCAACTTACTTCTGACAACGATCGGAGGACCGAAGGAG
CTAACCGCTTTTTTGCACAACATGGGGGATCATGTAACTCGCCTTGATCGTTGGGAACCG
GAGCTGAATGAAGCCATACCAAACGACGAGCGTGACACCACGATGCCTGTAGCAATGGCA
ACAACGTTGCGCAAACTATTAACTGGCGAACTACTTACTCTAGCTTCCCGGCAACAATTA
ATAGACTGGATGGAGGCGGATAAAGTTGCAGGACCACTTCTGCGCTCGGCCCTTCCGGCT
GGCTGGTTTATTGCTGATAAATCTGGAGCCGGTGAGCGTGGGTCTCGCGGTATCATTGCA
GCACTGGGGCCAGATGGTAAGCCCTCCCGTATCGTAGTTATCTACACGACGGGGAGTCAG
GCAACTATGGATGAACGAAATAGACAGATCGCTGAGATAGGTGCCTCACTGATTAAGCAT
TGGTAACTGTCAGACCAAGTTTACTCATATATACTTTAGATTGATTTAAAACTTCATTTT
TAATTTAAAAGGATCTAGGTGAAGATCCTTTTTGATAATCTCATGACCAAAATCCCTTAA
CGTGAGTTTTCGTTCCACTGAGCGTCAGACCCCGTAGAAAAGATCAAAGGATCTTCTTGA
GATCCTTTTTTTCTGCGCGTAATCTGCTGCTTGCAAACAAAAAAACCACCGCTACCAGCG
GTGGTTTGTTTGCCGGATCAAGAGCTACCAACTCTTTTTCCGAAGGTAACTGGCTTCAGC
AGAGCGCAGATACCAAATACTGTCCTTCTAGTGTAGCCGTAGTTAGGCCACCACTTCAAG
AACTCTGTAGCACCGCCTACATACCTCGCTCTGCTAATCCTGTTACCAGTGGCTGCTGCC
AGTGGCGATAAGTCGTGTCTTACCGGGTTGGACTCAAGACGATAGTTACCGGATAAGGCG
CAGCGGTCGGGCTGAACGGGGGGTTCGTGCACACAGCCCAGCTTGGAGCGAACGACCTAC
ACCGAACTGAGATACCTACAGCGTGAGCTATGAGAAAGCGCCACGCTTCCCGAAGGGAGA
AAGGCGGACAGGTATCCGGTAAGCGGCAGGGTCGGAACAGGAGAGCGCACGAGGGAGCTT
CCAGGGGGAAACGCCTGGTATCTTTATAGTCCTGTCGGGTTTCGCCACCTCTGACTTGAG
CGTCGATTTTTGTGATGCTCGTCAGGGGGGCGGAGCCTATGGAAAAACGCCAGCAACGCG
GCCTTTTTACGGTTCCTGGCCTTTTGCTGGCCTTTTGCTCACATGTTCTTTCCTGCGTTA
TCCCCTGATTCTGTGGATAACCGTATTACCGCCTTTGAGTGAGCTGATACCGCTCGCCGC
AGCCGAACGACCGAGCGCAGCGAGTCAGTGAGCGAGGAAGCGGAAGAGCGCCCAATACGC
AAACCGCCTCTCCCCGCGCGTTGGCCGATTCATTAATGCAGCTGGCACGACAGGTTTCCC
GACTGGAAAGCGGGCAGTGAGCGCAACGCAATTAATGTGAGTTAGCTCACTCATTAGGCA
CCCCAGGCTTTACACTTTATGCTTCCGGCTCGTATGTTGTGTGGAATTGTGAGCGGATAA
CAATTTCACACAGGAAACAGCTATGACCATGATTACGCCAAGCTCGAAATTAACCCTCAC
TAAAGGGAACAAAAGCTGGTACCCTTGTTTGTTAATGGTCAATTTAAGAATTATATAAAA
AGTTTTTTTTTTTTTGTAAATTATTAATAACTAATTTTTTTTTACAATTCGAACCAATTT
TTTAATTATTTCATGTTACTATATATCTGAGGATATAAATCAAAAAAATAAAATAAAAAG
TAGCACTGAAATAAAATGGAAGTAACGTCAACTTTATTAAAAAAAAGTCAAATGTTTGCA
GATGATTCATCTGATGGATTTCCTACAACAAAAAAATTTTTTGTTTCATCAATCGGTATT
ATATATTTTCATTTTGTTTTGAAAATTATAGGAATAAAATTAATTCACAATAACTTACTT
ATATGTTACAAGTTTATTTAATCGGTCTTTCCCACCTCATTCTTTTTAGGAAAGGCTCAT
TTAATAAATTCACTTTATGGAATTGGCTACACAATACAGATAGCTATGCTTCCTTACCTG
GTAAGTTTCATCATGTAAATACCACAAAATGATGGTTCGAAATATTTATTAGCTGTTTTT
TTTCACAAAAAATTACCAAATTAATACCTTAAAACAAAATAAATCACATTTAATCGTTAA
TATATATATGTGTAAATAATGGAAATACATAGACAATGGATAAATAAATACAATGGACGA
TCGATTCTATAAGGGCGAATTCTGCAGATATCCATCACACTGGCGGCCGCTCGAGCATGC
ATCTAGAGGGCC
Restriction sites to linearize plasmid
Partial or complete disruption of the genePartial
Additional remarks partial/complete disruption The promoter region and the first 2 exons of PB000201.02.0 remain intact
Selectable marker used to select the mutant parasitetgdhfr
Promoter of the selectable markerpbdhfr
Selection (positive) procedurepyrimethamine
Selection (negative) procedureNo
Additional remarks genetic modificationThe reverse primer of the 5' target sequence is methylated. The target region was therefore cloned KpnI/ApaI using the ApaI site of the TOPO vector in which it was subcloned after PCR amplification.
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 1CTTGTTTGTTAATGGTCAATTTAAGAATTATATAAAAAG
Additional information primer 1L2172 (KpnI); 5'
Sequence Primer 2ATAGAatcgatCGTCCATTGTATTTATTTATCCATTGTCTATG
Additional information primer 2L2173 (ClaI); 5' (ClaI site is Methylated!)
Sequence Primer 3ATAGAgatatcGCTGCTATGTGCTTTCATTTGTTTTG
Additional information primer 3L2174 (EcoRV); 3'
Sequence Primer 4ATAGAggatccGCAATTATTTACGAACAATTGATTCAGC
Additional information primer 4L2175 (BamHI); 3'
Sequence Primer 5
Additional information primer 5
Sequence Primer 6
Additional information primer 6