RMgmDB - Rodent Malaria genetically modified Parasites

Summary

RMgm-262
Malaria parasiteP. berghei
Genotype
DisruptedGene model (rodent): PBANKA_1018200; Gene model (P.falciparum): PF3D7_1426400; Gene product: MORN repeat protein, putative
PhenotypeNo phenotype has been described
Last modified: 19 March 2010, 08:34
  *RMgm-262
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
Name Group/DepartmentLeiden Malaria Research Group
Name InstituteLeiden University Medical Center
CityLeiden
CountryThe Netherlands
Name of the mutant parasite
RMgm numberRMgm-262
Principal name649cl2 (681 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 PBANKA_101820 (MORN repeat protein, putative)

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

Additional information

PBANKA_101820 (PF14_0243b) and PBANKA_101820 (PF14_0243a; dynein-associated protein, putative) were previously annotated as one gene.

Figure illustrating the PBANKA_101820 and PBANKA_101830 locus. Arrows indicate primer locations flanking the disrupted region of the targeted PBANKA_101820 locus.

Other mutants


  Disrupted: Mutant parasite with a disrupted gene
Details of the target gene
Gene Model of Rodent Parasite PBANKA_1018200
Gene Model P. falciparum ortholog PF3D7_1426400
Gene productMORN repeat 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
AAATTAAAAAAAAACATATAAACACAAATGATGTTTTTTCCTTCAATTTCGATATCGAAT
TCGAGAATATAAAGGAATATGGGAGAATGGGAAAAGTGTTAATGGAAATTATTATTTTAA
TGATGGTTTAAAATATGAAAATACATGGGATTATTTAACTGATTTTCCATATTTTTATAA
TGAAGAAATAAATAAAAATGAAATTATTTATCAAAAAGAAAAAAGAAATGAATATTTAGA
CAACATTTATGATATAGGTATAAAAAATTATAACTTTTTTTTTGCACTAATAAAAAAATA
CATATATATAGATACTATAGCAAAAATTTAAGTACTTTGTTTTCATCATTTTTTAGCTAA
ATATAATTTATATATGCGCAGTTTCATATCTATATAATAAACCGTTTTTTTTTTTTTTCC
TCTTATAACTAACATATTTTTTTTGTCAGACATATTCTTTATATAATCAAGAAATATAGT
TTTATTTCCTCTTCGAATGATAACAATAATCACCTTTTTTTTTAAGGTGATGGGTATTAT
ACAACCGATGGCGGCCGCCACCGCGGTGGAGCTCCAATTCGCCCTATAGTGAGTCGTATT
ACGCGCGCTCACTGGCCGTCGTTTTACAACGTCGTGACTGGGAAAACCCTGGCGTTACCC
AACTTAATCGCCTTGCAGCACATCCCCCTTTCGCCAGCTGGCGTAATAGCGAAGAGGCCC
GCACCGATCGCCCTTCCCAACAGTTGCGCAGCCTGAATGGCGAATGGGACGCGCCCTGTA
GCGGCGCATTAAGCGCGGCGGGTGTGGTGGTTACGCGCAGCGTGACCGCTACACTTGCCA
GCGCCCTAGCGCCCGCTCCTTTCGCTTTCTTCCCTTCCTTTCTCGCCACGTTCGCCGGCT
TTCCCCGTCAAGCTCTAAATCGGGGGCTCCCTTTAGGGTTCCGATTTAGTGCTTTACGGC
ACCTCGACCCCAAAAAACTTGATTAGGGTGATGGTTCACGTAGTGGGCCATCGCCCTGAT
AGACGGTTTTTCGCCCTTTGACGTTGGAGTCCACGTTCTTTAATAGTGGACTCTTGTTCC
AAACTGGAACAACACTCAACCCTATCTCGGTCTATTCTTTTGATTTATAAGGGATTTTGC
CGATTTCGGCCTATTGGTTAAAAAATGAGCTGATTTAACAAAAATTTAACGCGAATTTTA
ACAAAATATTAACGCTTACAATTTAGGTGGCACTTTTCGGGGAAATGTGCGCGGAACCCC
TATTTGTTTATTTTTCTAAATACATTCAAATATGTATCCGCTCATGAGACAATAACCCTG
ATAAATGCTTCAATAATATTGAAAAAGGAAGAGTATGAGTATTCAACATTTCCGTGTCGC
CCTTATTCCCTTTTTTGCGGCATTTTGCCTTCCTGTTTTTGCTCACCCAGAAACGCTGGT
GAAAGTAAAAGATGCTGAAGATCAGTTGGGTGCACGAGTGGGTTACATCGAACTGGATCT
CAACAGCGGTAAGATCCTTGAGAGTTTTCGCCCCGAAGAACGTTTTCCAATGATGAGCAC
TTTTAAAGTTCTGCTATGTGGCGCGGTATTATCCCGTATTGACGCCGGGCAAGAGCAACT
CGGTCGCCGCATACACTATTCTCAGAATGACTTGGTTGAGTACTCACCAGTCACAGAAAA
GCATCTTACGGATGGCATGACAGTAAGAGAATTATGCAGTGCTGCCATAACCATGAGTGA
TAACACTGCGGCCAACTTACTTCTGACAACGATCGGAGGACCGAAGGAGCTAACCGCTTT
TTTGCACAACATGGGGGATCATGTAACTCGCCTTGATCGTTGGGAACCGGAGCTGAATGA
AGCCATACCAAACGACGAGCGTGACACCACGATGCCTGTAGCAATGGCAACAACGTTGCG
CAAACTATTAACTGGCGAACTACTTACTCTAGCTTCCCGGCAACAATTAATAGACTGGAT
GGAGGCGGATAAAGTTGCAGGACCACTTCTGCGCTCGGCCCTTCCGGCTGGCTGGTTTAT
TGCTGATAAATCTGGAGCCGGTGAGCGTGGGTCTCGCGGTATCATTGCAGCACTGGGGCC
AGATGGTAAGCCCTCCCGTATCGTAGTTATCTACACGACGGGGAGTCAGGCAACTATGGA
TGAACGAAATAGACAGATCGCTGAGATAGGTGCCTCACTGATTAAGCATTGGTAACTGTC
AGACCAAGTTTACTCATATATACTTTAGATTGATTTAAAACTTCATTTTTAATTTAAAAG
GATCTAGGTGAAGATCCTTTTTGATAATCTCATGACCAAAATCCCTTAACGTGAGTTTTC
GTTCCACTGAGCGTCAGACCCCGTAGAAAAGATCAAAGGATCTTCTTGAGATCCTTTTTT
TCTGCGCGTAATCTGCTGCTTGCAAACAAAAAAACCACCGCTACCAGCGGTGGTTTGTTT
GCCGGATCAAGAGCTACCAACTCTTTTTCCGAAGGTAACTGGCTTCAGCAGAGCGCAGAT
ACCAAATACTGTCCTTCTAGTGTAGCCGTAGTTAGGCCACCACTTCAAGAACTCTGTAGC
ACCGCCTACATACCTCGCTCTGCTAATCCTGTTACCAGTGGCTGCTGCCAGTGGCGATAA
GTCGTGTCTTACCGGGTTGGACTCAAGACGATAGTTACCGGATAAGGCGCAGCGGTCGGG
CTGAACGGGGGGTTCGTGCACACAGCCCAGCTTGGAGCGAACGACCTACACCGAACTGAG
ATACCTACAGCGTGAGCTATGAGAAAGCGCCACGCTTCCCGAAGGGAGAAAGGCGGACAG
GTATCCGGTAAGCGGCAGGGTCGGAACAGGAGAGCGCACGAGGGAGCTTCCAGGGGGAAA
CGCCTGGTATCTTTATAGTCCTGTCGGGTTTCGCCACCTCTGACTTGAGCGTCGATTTTT
GTGATGCTCGTCAGGGGGGCGGAGCCTATGGAAAAACGCCAGCAACGCGGCCTTTTTACG
GTTCCTGGCCTTTTGCTGGCCTTTTGCTCACATGTTCTTTCCTGCGTTATCCCCTGATTC
TGTGGATAACCGTATTACCGCCTTTGAGTGAGCTGATACCGCTCGCCGCAGCCGAACGAC
CGAGCGCAGCGAGTCAGTGAGCGAGGAAGCGGAAGAGCGCCCAATACGCAAACCGCCTCT
CCCCGCGCGTTGGCCGATTCATTAATGCAGCTGGCACGACAGGTTTCCCGACTGGAAAGC
GGGCAGTGAGCGCAACGCAATTAATGTGAGTTAGCTCACTCATTAGGCACCCCAGGCTTT
ACACTTTATGCTTCCGGCTCGTATGTTGTGTGGAATTGTGAGCGGATAACAATTTCACAC
AGGAAACAGCTATGACCATGATTACGCCAAGCTCGAAATTAACCCTCACTAAAGGGAACA
AAAGCTGGTACCCTACTTTGTTATAAAGACATGTACTTATATTTCTCTTAGATGGTCAAA
CGCCTTGAAAATGAAATAAATAAATCAATCAATAAATAAATAATTAATAAATAAGACAAT
GTAAAATCATTGGCGCAAAAAAAATATATTATAATTAAAAATGTTATTTGCCTATCCAAC
TCGATAAAAAAAAATATTTGCGAACTCATTGTTTAATAAATTGCTTACATGGGCAATATA
TTTATGTCTAAGTATGCATTCCATGATAAAATTAACTATCTCCTTATTACATGGAGCCTA
TTATTTTGCATGTTTATTCATTCATTCATTCCGTTATTTGTTTATTCATTTTTTTTATTT
ATTTTTCAAACTTTACCCACAAACTCATATACAAAAGCCATTAACATATTCCATTTATGA
AAATGTTAAAATTACCTCGAAATATATTTCTAACATTTTAACATTTTTACTAAATCATCG
TTTCAGTTCCTCTGAGGTATAAAACAAAATATTCAGTTACGCAGATACATGTAAATATGT
GTGCATGTGTGAAAAAAGCTATCATAAAATTTTTCCAATTTAAGCTAGCTCGTAAATATT
TTTTTTTTCACACACAGTATTAAACGCTAGCATCGTTCGTTTATTTTTTCATTTCGTTGT
TTTGCCATTTCGTAGTTTTATTATTCTACTTATTTCGTTTATTTACATAAAACAGTATCC
TATGTCGATCGATTCTATAAGGGCGAATTCTGCAGATATCCATCACACTGGCGGCCGCTC
GAGCATGCATCTAGAGGGCC
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 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 1ATAGAggtaccCTACTTTGTTATAAAGACATGTACTTATATTTCTCTTAG
Additional information primer 1L2176 (KpnI); 5'
Sequence Primer 2ATAGAatcgatCGACATAGGATACTGTTTTATGTAAATAAACG
Additional information primer 2L2177 (ClaI); 5' (ClaI site Methylated!)
Sequence Primer 3ATAGAgaattcGAGAATATAAAGGAATATGGGAGAATGG
Additional information primer 3L2178 (EcoRI); 3'
Sequence Primer 4ATAGAgcggccgcCATCGGTTGTATAATACCCATCACC
Additional information primer 4L2179 (NotI); 3'
Sequence Primer 5
Additional information primer 5
Sequence Primer 6
Additional information primer 6