RMgmDB - Rodent Malaria genetically modified Parasites

Summary

RMgm-882
Malaria parasiteP. berghei
Genotype
Genetic modification not successful
DisruptedGene model (rodent): PBANKA_1222500; Gene model (P.falciparum): PF3D7_0808200; Gene product: plasmepsin X
PhenotypeNo phenotype has been described
Last modified: 8 June 2014, 12:30
  *RMgm-882
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) 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 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. This reference line does not contain a drug-selectable marker (PubMed: PMID: 16242190)
Attempts to generate the mutant parasite were performed by
Name PI/ResearcherB. Franke-Fayard; J.B. Dame; C.J. Janse
Name Group/DepartmentLeiden Malaria Research Group
Name InstituteLeiden University Medical Center (LUMC)
CityLeiden
CountryThe Netherlands

  Disrupted: Mutant parasite with a disrupted gene
Details of the target gene
Gene Model of Rodent Parasite PBANKA_1222500
Gene Model P. falciparum ortholog PF3D7_0808200
Gene productplasmepsin X
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
Click to view information
Click to hide information
Plasmid/construct sequence
Click to view information
Click to hide information
ATGAAGAGCATAAAAATATTACCCGTATTTTATTTGGTGACATTTTTTTTGCACAACTAT
AATGAGATAAAATGTAACAAAGTATACATATCTGGAGATAATTTTATGCCTTGTTCAGAA
TGTAAAGATATAAATAAGTGTGGTGGTTGTTTGTTTGATAACAATGAGTTTTTACCTAGT
GCAATAGAATTAAAATTGACGAAAAAAAACAGGGATAACAATAATCATGATAATTTAAAA
ATTCACCATGATTCATTAAAATTAGGAAATGTAAAATATTATGTAAAAAGAGGAGAAGGT
GTTTCAGGAAGTTTTGGGAATGCTTCAGGAAATGACATAAACAGTATGGCTAAGATTCAT
AATGAAATAAAAAATCGAACAGAAAATAAGGAAGAAAATAAATCGTCTTTAAGTTTCATT
GATCATATTAATAATAATCTGGGAGAAGGAAAAGGAGGTTATAATTTTTCAAGAATACAA
AAACATGAACAGGATGGAGATAAAATAAATGCACAAGAAGAATTTGAGAAAATAAAAAGT
CAAGTTGTTGATAAATCTGTAGTTTCATTCTCTGATCGTGTTTTAGATGAAAATGAAGGT
CAAACTCAGCCTTCCAAAGGTGTGACTGTTGAAGAAACAAGTGATAATGTGTTTTTAGTT
CCTTTACAACATTTAAGAGATAGTCAGTTTGTTGGTAAGTTATTAGTAGGTACACCTCCT
CAAGAAATTCATCCAATAAGCTTGCATGCCTGCAGGTCAACAATAAATAATAAATAAATA
TTGTGGAAATAAAATAACATATAATTATTTTTAATACATTGATTTCCCTTTTATTTTTTT
AAATTTCATTGATATAAAAATATATAATAATAACATATATGATTTCAAATTAATCTTTTC
AAAAATGGTGTTTATTTTTGTATGTTTGTGTATGAATTAATCACATAACACATCTATTAA
ATTGAGTTGGTAATATAGACACAAATAAATATATATATTTTTATAGCTTAAAAGTGTGTT
ATGAATATTTTAAGCATATTTTCTTTTTCTTTGGATTGTGTAAAATGAACTCATATAATG
CGTTTTTTTGTTTTTGTTATTTTGTCATTTTGTTATTTTGCTATTTTATGGATTAATTTT
TGTTTATAAAATGGGAAATAATTTTAACATATTTAAATAAATGGAGAAAAAATATAAAAT
AATTATAAAAAAAAGTTAATACACATTTTTTCCTGTTATAGACCTTATATTTATTTATCC
ATATATATATATATATATATATATATATATATATACATACCAAGTGAATTAAGAGGAAAG
CTAATTTATTATTCAGAATAATATATGAACTATATATAATTTTTATTATTTTGGTGTATA
TTAATCTGTCTATATGCATACATGCAATAATTTATCGACTTATATATCAAATAACATAAA
ATAGAAGTGTTTTAAATTATGGATATATGCTCAATATTCATTTTTTTTAATAAGTTAGCT
ATATTTAAATTATACATTTTATATATGGTCTCTTTTTTTTTTAAATATTATTTAAGTGAT
CATGAAAATATAAATAATTTTTTTTTATTTAATATCCTTTTGCTTGCATGTGGTAAATGG
AAATTTGGATGTGTTTTGAARGTTCGGATATAGTTGTATGGACATATAATATATTTTGTG
AAAAATTGGTTTTATGTTTATACTTATGCCAATACTTTTTGAGTAAAACAAAGCAAGTGC
TTATAAATAATTAAAGCCAATTTTATAATATATATTTTTTTATTTAATTTGAATTTAGTA
GTATAATTTTTTATGGTAAGTGCTCAAAGAGAGTTGCTTATAAAGTATGGTTTGTTTCTT
TTTCGCCATTTTGAATTACACATTAAAAATATATAGATACATATATTATAATATGAAATC
ATTAATAATTTAGGGAAATTCTACAAATTTAAAAACGAATAAAATAATTGTTTTTCATCA
TGCCATAACACAATATTGATATATACATGTACAAACATTTTTTTTATTTGGAAAATATAA
ATTATATAAAAAAAAATGTATAGTATACAAAATGAGCATATTCACACGGGGTGGACGTTC
ATTTTTTCATTTTTCCCCTGTTTTTTATGAGTATATGATAAAATTTTATGAACATTTACA
CAAAATGAAAATGGATATATAGGAAAAATGGAGCGGTATTTCATTTATCTTTGATTGTCA
TTTGGATATTATATTACCYTGGGTAGGCAATTAAAAATGTTAAATAACAATTTAAGGAAA
TTATATTTTATATATTAAAATTAACACTGTATTATATGATTCGCTTATAAAAGCCACTCT
TTCCCCATGCAAAGCTGTTTAATATCAATTTTAACAAATTACACACATGTTAATATATTT
ATATATATAATTTATATATTTATAATTTATATATTTATATTTTTATTATTTATATATTTA
TTATTTATTGTGTGTGTCAATTCGGGTAGGATATACCTCTTTTTTATTGTTTAAAGCGAT
TTGTATTCTAAAATATAAAGRATTTGAAAAAGAGAAAGATAGAATATGATCCCATCATAT
ATAGCCCTATAATTTTTATTTAGCAGCGAATTAATTTTTCTATTAAGTTTATGTGTAATT
AAAATAACGGAATATATATAATACAATAAAAAAGTGCATAAATTAAAATTTTTTCAATTA
AATTTTTTTTTTTAAGGGGTTATATAATATTAAATATATAAAATACGATTATATATTTTT
GCTACAATTTTTTATATTAAGATATAAATAGTAAATAAATGGTATTATATGGCATGTAAT
ATATAAATTTTTTCCAATTTTTATTTTATATACACTTTTCCTTTTTTTGTCATAAAACTT
AAACAATTTACACATTCATTTTAAAAATTGACTATTTGTTTCAACATTTTTTGAGTTTCC
GTTTTATAATAGTATTTTCATTTGTATATTGCTTATATATATAAATACACACCTAAATGT
TACAAAGGATCAATGCATAAACCGGTGTGTCTGGTCGTCGCGATGACCCCCAAGAGGGGC
ATCGGCATCAACAACGGCCTCCCGTGGCCCCACTTGACCACAGATTTCAAACACTTTTCT
CGTGTGACAAAAACGACGCCCGAAGAAGCCAGTCGCCTGAACGGGTGGCTTCCCAGGAAA
TTTGCAAAGACGGGCGACTCTGGACTTCCCTCTCCATCAGTCGGCAAGAGATTCAACGCC
GTTGTCATGGGACGGAAAACCTGGGAAAGCATGCCTCGAAAGTTTAGACCCCTCGTGGAC
AGATTGAACATCGTCGTTTCCTCTTCCCTCAAAGAAGAAGACATTGCGGCGGAGAAGCCT
CAAGCTGAAGGCCAGCAGCGCGTCCGAGTCTGTGCTTCACTCCCAGCAGCTCTCAGCCTT
CTGGAGGAAGAGTACAAGGATTCTGTCGACCAGATTTTTGTCGTGGGAGGAGCGGGACTG
TACGAGGCAGCGCTGTCTCTGGGCGTTGCCTCTCACCTGTACATCACGCGTGTAGCCCGC
GAGTTTCCGTGCGACGTTTTCTTCCCTGCGTTCCCCGGAGATGACATTCTTTCAAACAAA
TCAACTGCTGCGCAGGCTGCAGCTCCTGCCGAGTCTGTGTTCGTTCCCTTTTGTCCGGAG
CTCGGAAGAGAGAAGGACAATGAAGCGACGTATCGACCCATCTTCATTTCCAAGACCTTC
TCAGACAACGGGGTTCCCTACGACTTTGTGGTTCTCGAGAAGAGAAGGAAGACTGACGAC
GCAGCCACTGCGGAACCGAGCAACGCAATGAGCTCCTTGACGTCCACGAGGGAGACAACT
CCCGTGCACGGGTTGCAGGCTCCTTCTTCGGCCGCAGCCATTGCCCCGGTGTTGGCGTGG
ATGGACGAAGAAGACCGGAAAAAACGCGAGCAAAAGGAACTGATTCGGGCCGTTCCGCAT
GTTCACTTTAGAGGCCATGAAGAGTTCCAGTACCTTGATCTCATTGCCGACATTATTAAC
AATGGAAGGACAATGGATGACCGAACGGGCGTTGGTGTCATCTCCAAATTCGGCTGCACT
ATGCGCTACTCGCTGGATCAGGCCTTTCCACTTCTCACCACAAAGCGTGTGTTCTGGAAA
GGGGTCCTCGAAGAGTTGCTGTGGTTCATTCGCGGCGACACGAACGCAAACCATCTTTCT
GAGAAGGGCGTGAAGATCTGGGACAAGAATGTGACACGCGAGTTCCTCGATTCGCGCAAT
CTCCCCCACCGAGAGGTCGGAGACATCGGCCCGGGCTACGGCTTCCAGTGGAGACACTTC
GGCGCGGCATACAAAGACATGCACACAGACTACACAGGGCAGGGCGTCGACCAGCTGAAG
AATGTGATCCAGATGCTGAGAACGAATCCAACAGATCGTCGCATGCTCATGACTGCCTGG
AATCCTGCAGCGCTGGACGAAATGGCGCTGCCGCCTTGTCACTTGTTGTGCCAGTTCTAC
GTGAACGACCAGAAGGAGCTGTCGTGCATCATGTATCAGCGGTCGTGCGATGTCGGCCTC
GGCGTCCCCTTCAACATCGCTTCCTATTCGCTTTTGACGCTCATGGTTGCACACGTCTGC
AACCTAAAACCTAAGGAGTTCATTCACTTCATGGGGAACACGCATGTCTACACGAACCAT
GTCGAGGCTTTAAAAGAGCAGCTGCGGAGAGAACCGAGACCGTTCCCCATTGTGAACATC
CTCAACAAGGAACGCATCAAGGAAATCGACGATTTCACCGCCGAGGATTTTGAGGTCGTG
GGCTACGTCCCGCACGGACGAATCCAGATGGAGATGGCTGTCTAGCGGAAATACAGAAGC
TAGCTTTGATCCCGTTTTTCTTACTTATATATTTATACCAATTGATTGTATTTATAACTG
TAAAAATGTGTATGTTGTGTGCATATTTTTTTTTGTGCATGCACATGCATGTAAATAGCT
AAAATTATGAACATTTTATTTTTTGTTCAGAAAAAAAAAACTTTACACACATAAAATGGC
TAGTATGAATAGCCATATTTTATATAAATTAAATCCTATGAATTTATGACCATATTAAAA
ATTTAGATATTTATGGAACATAATATGTTTGAAACAATAAGACAAAATTATTATTATTAT
TATTATTTTTACTGTTATAATTATGTTGTCTCTTCAATGATTCATAAATAGTTGGACTTG
ATTTTTAAAATGTTTATAATATGATTAGCATAGTTAAATAAAAAAAGTTGAAAAATTAAA
AAAAAACATATAAACACAAATGATGTTTTTTCCTTCAATTTCGATATCGAATTCCTGCAG
CCCGGGGGATCCTGGTATGCTAACTGCAGGAAATATTCCATTTTTCGATAATTTATTAAA
ACAATATAAAAATATGACACCTCAATTTTCTTTTTATATATCTCCAAATGATGAAACATC
TACATTTATAGTTGGTGGAATAAGCAAATCATATTATGAAGGAGATATATATATGTTACC
TGTTGTTAAAGAATATTATTGGGAAGTTAAGTTAGATGCTATATATATAGGAGATGAAAA
AATATGTTGCGAAGAGGAAAGTTATGCAATATTTGATTCAGGAACTTCTTACAATACAAT
GCCTAGTACTCAAATAAGTAATTTTTTTAAGATTGTTTCATCAAAACCATGTAATGAAGA
AAATTATAACAATATATTAAAAGAATATCCAACTATAAAATATGTTTTTGGAAAGTTAGT
AATTGAACTATTACCCAATGAATATATGATTGTAAATGACGATTTATGTGTTCCAGCTTA
TATGCAGATCGATGTTCCATCAGAAAATAATAATGCTTATCTTCTTGGAACTATAGCTTT
TATGAGACATTACTTTACAGTTTTTGTAAGAGGGCAAGAAGGGAATCCATCTATGGTTGG
GGTGGCAAAAGCTAAACAGGTTTAATTTTGCTTTAGAATATAGTAAATGAATATTATGGG
AGAGGAATTCATAAATAGAAACGAATTGAAAATACGCAGAAACAAAAACGAATTGAGATG
TAATGAGAATTACAGGGCGGCCGCCACCGCGGTGGAGCTCCAATTCGCCCTATAGTGAGT
CGTATTACAATTCACTGGCCGTCGTTTTACAACGTCGTGACTGGGAAAACCCTGGCGTTA
CCCAACTTAATCGCCTTGCAGCACATCCCCCTTTCGCCAGCTGGCGTAATAGCGAAGAGG
CCCGCACCGATCGCCCTTCCCAACAGTTGCGCAGCCTGAATGGCGAATGGGACGCGCCCT
GTAGCGGCGCATTAAGCGCGGCGGGTGTGGTGGTTACGCGCAGCGTGACCGCTACACTTG
CCAGCGCCCTAGCGCCCGCTCCTTTCGCTTTCTTCCCTTCCTTTCTCGCCACGTTCGCCG
GCTTTCCCCGTCAAGCTCTAAATCGGGGGCTCCCTTTAGGGTTCCGATTTAGTGCTTTAC
GGCACCTCGACCCCAAAAAACTTGATTAGGGTGATGGTTCACGTAGTGGGCCATCGCCCT
GATAGACGGTTTTTCGCCCTTTGACGTTGGAGTCCACGTTCTTTAATAGTGGACTCTTGT
TCCAAACTGGAACAACACTCAACCCTATCTCGGTCTATTCTTTTGATTTATAAGGGATTT
TGCCGATTTCGGCCTATTGGTTAAAAAATGAGCTGATTTAACAAAAATTTAACGCGAATT
TTAACAAAATATTAACGCTTACAATTTAGGTGGCACTTTTCGGGGAAATGTGCGCGGAAC
CCCTATTTGTTTATTTTTCTAAATACATTCAAATATGTATCCGCTCATGAGACAATAACC
CTGATAAATGCTTCAATAATATTGAAAAAGGAAGAGTATGAGTATTCAACATTTCCGTGT
CGCCCTTATTCCCTTTTTTGCGGCATTTTGCCTTCCTGTTTTTGCTCACCCAGAAACGCT
GGTGAAAGTAAAAGATGCTGAAGATCAGTTGGGTGCACGAGTGGGTTACATCGAACTGGA
TCTCAACAGCGGTAAGATCCTTGAGAGTTTTCGCCCCGAAGAACGTTTTCCAATGATGAG
CACTTTTAAAGTTCTGCTATGTGGCGCGGTATTATCCCGTATTGACGCCGGGCAAGAGCA
ACTCGGTCGCCGCATACACTATTCTCAGAATGACTTGGTTGAGTACTCACCAGTCACAGA
AAAGCATCTTACGGATGGCATGACAGTAAGAGAATTATGCAGTGCTGCCATAACCATGAG
TGATAACACTGCGGCCAACTTACTTCTGACAACGATCGGAGGACCGAAGGAGCTAACCGC
TTTTTTGCACAACATGGGGGATCATGTAACTCGCCTTGATCGTTGGGAACCGGAGCTGAA
TGAAGCCATACCAAACGACGAGCGTGACACCACGATGCCTGTAGCAATGGCAACAACGTT
GCGCAAACTATTAACTGGCGAACTACTTACTCTAGCTTCCCGGCAACAATTAATAGACTG
GATGGAGGCGGATAAAGTTGCAGGACCACTTCTGCGCTCGGCCCTTCCGGCTGGCTGGTT
TATTGCTGATAAATCTGGAGCCGGTGAGCGTGGGTCTCGCGGTATCATTGCAGCACTGGG
GCCAGATGGTAAGCCCTCCCGTATCGTAGTTATCTACACGACGGGGAGTCAGGCAACTAT
GGATGAACGAAATAGACAGATCGCTGAGATAGGTGCCTCACTGATTAAGCATTGGTAACT
GTCAGACCAAGTTTACTCATATATACTTTAGATTGATTTAAAACTTCATTTTTAATTTAA
AAGGATCTAGGTGAAGATCCTTTTTGATAATCTCATGACCAAAATCCCTTAACGTGAGTT
TTCGTTCCACTGAGCGTCAGACCCCGTAGAAAAGATCAAAGGATCTTCTTGAGATCCTTT
TTTTCTGCGCGTAATCTGCTGCTTGCAAACAAAAAAACCACCGCTACCAGCGGTGGTTTG
TTTGCCGGATCAAGAGCTACCAACTCTTTTTCCGAAGGTAACTGGCTTCAGCAGAGCGCA
GATACCAAATACTGTCCTTCTAGTGTAGCCGTAGTTAGGCCACCACTTCAAGAACTCTGT
AGCACCGCCTACATACCTCGCTCTGCTAATCCTGTTACCAGTGGCTGCTGCCAGTGGCGA
TAAGTCGTGTCTTACCGGGTTGGACTCAAGACGATAGTTACCGGATAAGGCGCAGCGGTC
GGGCTGAACGGGGGGTTCGTGCACACAGCCCAGCTTGGAGCGAACGACCTACACCGAACT
GAGATACCTACAGCGTGAGCTATGAGAAAGCGCCACGCTTCCCGAAGGGAGAAAGGCGGA
CAGGTATCCGGTAAGCGGCAGGGTCGGAACAGGAGAGCGCACGAGGGAGCTTCCAGGGGG
AAACGCCTGGTATCTTTATAGTCCTGTCGGGTTTCGCCACCTCTGACTTGAGCGTCGATT
TTTGTGATGCTCGTCAGGGGGGCGGAGCCTATGGAAAAACGCCAGCAACGCGGCCTTTTT
ACGGTTCCTGGCCTTTTGCTGGCCTTTTGCTCACATGTTCTTTCCTGCGTTATCCCCTGA
TTCTGTGGATAACCGTATTACCGCCTTTGAGTGAGCTGATACCGCTCGCCGCAGCCGAAC
GACCGAGCGCAGCGAGTCAGTGAGCGAGGAAGCGGAAGAGCGCCCAATACGCAAACCGCC
TCTCCCCGCGCGTTGGCCGATTCATTAATGCAGCTGGCACGACAGGTTTCCCGACTGGAA
AGCGGGCAGTGAGCGCAACGCAATTAATGTGAGTTAGCTCACTCATTAGGCACCCCAGGC
TTTACACTTTATGCTTCCGGCTCGTATGTTGTGTGGAATTGTGAGCGGATAACAATTTCA
CACAGGAAACAGCTATGACCATGATTACGCCAAGCTCGAAATTAACCCTCACTAAAGGGA
ACAAAAGCTGGTACC
Restriction sites to linearize plasmid
Partial or complete disruption of the geneUnknown
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 negative attempts to disrupt plasmepsin X indicates an essential role during blood stage development.

See also RMgm-84 for an independent, unsuccessful attempt to disrupt plasmepsin X

P. berghei has 7 plasmepsins (aspartic proteases (PM IV-PM X). PM IV has a role in hemoglobin digestion (RMgm-314, RMgm-315, RMgm-316). P. falciparum PM V performs a critic role in the endoplasmic reticulum processing the PEXEL sequence of exported proteins. PM VI, VII, VII are not expressed in asexual blood stages; these are expressed in gametocytes/ookinetes/oocysts. PM VI plays an essential role in the formation of sporozoites within the oocyst (RMgm-97).
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 1
Additional information primer 1
Sequence Primer 2
Additional information primer 2
Sequence Primer 3
Additional information primer 3
Sequence Primer 4
Additional information primer 4
Sequence Primer 5
Additional information primer 5
Sequence Primer 6
Additional information primer 6