Summary

RMgm-108
Malaria parasiteP. berghei
Genotype
DisruptedGene model (rodent): PBANKA_1309600; Gene model (P.falciparum): PF3D7_1445800; Gene product: conserved Plasmodium membrane protein, unknown function
Phenotype Oocyst; Sporozoite;
Last modified: 18 February 2009, 11:56
  *RMgm-108
Successful modificationThe parasite was generated by the genetic modification
The mutant contains the following genetic modification(s) Gene disruption
Reference (PubMed-PMID number) Reference 1 (PMID number) : 18974882
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 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).
The mutant parasite was generated 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
Name of the mutant parasite
RMgm numberRMgm-108
Principal name802cl1; 838
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
OocystOocyst numbers in A. stephensi were comparable to wild type (ranging from 150–250 oocysts per mosquito). Sporozoite formation blocked inside the oocysts. No sporozoite formation was detectable within the oocysts by either fluorescence or phase-contrast microscopy.
SporozoiteSporozoite formation blocked inside the oocysts. No sporozoite formation was detectable within the oocysts by either fluorescence or phase-contrast microscopy. No sporozoites in salivary glands. No blood stage infection in C57BL/6 mice after feeding of infected mosquitoes on these mice.
Liver stageNot tested
Additional remarks phenotype

Mutant/mutation
The mutant lacks expression of PB101363.00.0

Protein (function)
Unknown. The protein was detected in a proteome analysis of P. falciparum oocysts and sporozoites. It was highly abundant in oocyst-derived sporozoites and not detected in salivary gland sporozoites. It contains a signal peptide and a transmembrane region.

Phenotype
The phenotype analyses indicate a role of the protein in the formation of sporozoites within the oocysts. The mutant produces normal numbers of oocysts; however,  no sporozoite formation was detectable within these oocysts.This early function in sporozoite development of this protein is in agreement with its presence in oocyst derived sporozoites and absence in salivary gland sporozoites.


  Disrupted: Mutant parasite with a disrupted gene
Details of the target gene
Gene Model of Rodent Parasite PBANKA_1309600
Gene Model P. falciparum ortholog PF3D7_1445800
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
ACCCTCACTAAAGGGAACAAAAGCTGGTACCAAATAGTGCCTTTTTCGGTTTTTATGTTC
TCAAATACAATTTCATAGGTTTAATGTTGGTAGACAGGTAATTTTATAGTCAATATAATT
TAAAGCTAATATAATTTGTATAATATTTAACATAGGAACTTTTAAAAATATAATTATATT
TTTTTATACAATTTTTTAAATATATAAATCGTTGAAAAAGTATTGGAAAATAAAGATTGT
ATATATATACGCATGCACCCATGTAACTATAATGCAAGCACAAACGTTTATTCAATCTAA
AATATACATATATATATTGATATAAAAAAAAATAAAAATAAGCCCTATATTTCAAATTAC
TACTAAAATAAAGATATATCTTGGAAGCATAAATATATTATTTCAAGAAATATTTTTAGA
AGTCTTATAACCATTCTCAAGCTTGCATGCCTGCAGGTCAACAATAAATAATAAATAAAT
ATTGTGGAAATAAAATAACATATAATTATTTTTAATACATTGATTTCCCTTTTATTTTTT
TAAATTTCATTGATATAAAAATATATAATAATAACATATATGATTTCAAATTAATCTTTT
CAAAAATGGTGTTTATTTTTGTATGTTTGTGTATGAATTAATCACATAACACATCTATTA
AATTGAGTTGGTAATATAGACACAAATAAATATATATATTTTTATAGCTTAAAAGTGTGT
TATGAATATTTTAAGCATATTTTCTTTTTCTTTGGATTGTGTAAAATGAACTCATATAAT
GCGTTTTTTTGTTTTTGTTATTTTGTCATTTTGTTATTTTGCTATTTTATGGATTAATTT
TTGTTTATAAAATGGGAAATAATTTTAACATATTTAAATAAATGGAGAAAAAATATAAAA
TAATTATAAAAAAAAGTTAATACACATTTTTTCCTGTTATAGACCTTATATTTATTTATC
CATATATATATATATATATATATATATATATATATACATACCAAGTGAATTAAGAGGAAA
GCTAATTTATTATTCAGAATAATATATGAACTATATATAATTTTTATTATTTTGGTGTAT
ATTAATCTGTCTATATGCATACATGCAATAATTTATCGACTTATATATCAAATAACATAA
AATAGAAGTGTTTTAAATTATGGATATATGCTCAATATTCATTTTTTTTAATAAGTTAGC
TATATTTAAATTATACATTTTATATATGGTCTCTTTTTTTTTTAAATATTATTTAAGTGA
TCATGAAAATATAAATAATTTTTTTTTATTTAATATCCTTTTGCTTGCATGTGGTAAATG
GAAATTTGGATGTGTTTTGAARGTTCGGATATAGTTGTATGGACATATAATATATTTTGT
GAAAAATTGGTTTTATGTTTATACTTATGCCAATACTTTTTGAGTAAAACAAAGCAAGTG
CTTATAAATAATTAAAGCCAATTTTATAATATATATTTTTTTATTTAATTTGAATTTAGT
AGTATAATTTTTTATGGTAAGTGCTCAAAGAGAGTTGCTTATAAAGTATGGTTTGTTTCT
TTTTCGCCATTTTGAATTACACATTAAAAATATATAGATACATATATTATAATATGAAAT
CATTAATAATTTAGGGAAATTCTACAAATTTAAAAACGAATAAAATAATTGTTTTTCATC
ATGCCATAACACAATATTGATATATACATGTACAAACATTTTTTTTATTTGGAAAATATA
AATTATATAAAAAAAAATGTATAGTATACAAAATGAGCATATTCACACGGGGTGGACGTT
CATTTTTTCATTTTTCCCCTGTTTTTTATGAGTATATGATAAAATTTTATGAACATTTAC
ACAAAATGAAAATGGATATATAGGAAAAATGGAGCGGTATTTCATTTATCTTTGATTGTC
ATTTGGATATTATATTACCYTGGGTAGGCAATTAAAAATGTTAAATAACAATTTAAGGAA
ATTATATTTTATATATTAAAATTAACACTGTATTATATGATTCGCTTATAAAAGCCACTC
TTTCCCCATGCAAAGCTGTTTAATATCAATTTTAACAAATTACACACATGTTAATATATT
TATATATATAATTTATATATTTATAATTTATATATTTATATTTTTATTATTTATATATTT
ATTATTTATTGTGTGTGTCAATTCGGGTAGGATATACCTCTTTTTTATTGTTTAAAGCGA
TTTGTATTCTAAAATATAAAGRATTTGAAAAAGAGAAAGATAGAATATGATCCCATCATA
TATAGCCCTATAATTTTTATTTAGCAGCGAATTAATTTTTCTATTAAGTTTATGTGTAAT
TAAAATAACGGAATATATATAATACAATAAAAAAGTGCATAAATTAAAATTTTTTCAATT
AAATTTTTTTTTTTAAGGGGTTATATAATATTAAATATATAAAATACGATTATATATTTT
TGCTACAATTTTTTATATTAAGATATAAATAGTAAATAAATGGTATTATATGGCATGTAA
TATATAAATTTTTTCCAATTTTTATTTTATATACACTTTTCCTTTTTTTGTCATAAAACT
TAAACAATTTACACATTCATTTTAAAAATTGACTATTTGTTTCAACATTTTTTGAGTTTC
CGTTTTATAATAGTATTTTCATTTGTATATTGCTTATATATATAAATACACACCTAAATG
TTACAAAGGATCAATGCATAAACCGGTGTGTCTGGTCGTCGCGATGACCCCCAAGAGGGG
CATCGGCATCAACAACGGCCTCCCGTGGCCCCACTTGACCACAGATTTCAAACACTTTTC
TCGTGTGACAAAAACGACGCCCGAAGAAGCCAGTCGCCTGAACGGGTGGCTTCCCAGGAA
ATTTGCAAAGACGGGCGACTCTGGACTTCCCTCTCCATCAGTCGGCAAGAGATTCAACGC
CGTTGTCATGGGACGGAAAACCTGGGAAAGCATGCCTCGAAAGTTTAGACCCCTCGTGGA
CAGATTGAACATCGTCGTTTCCTCTTCCCTCAAAGAAGAAGACATTGCGGCGGAGAAGCC
TCAAGCTGAAGGCCAGCAGCGCGTCCGAGTCTGTGCTTCACTCCCAGCAGCTCTCAGCCT
TCTGGAGGAAGAGTACAAGGATTCTGTCGACCAGATTTTTGTCGTGGGAGGAGCGGGACT
GTACGAGGCAGCGCTGTCTCTGGGCGTTGCCTCTCACCTGTACATCACGCGTGTAGCCCG
CGAGTTTCCGTGCGACGTTTTCTTCCCTGCGTTCCCCGGAGATGACATTCTTTCAAACAA
ATCAACTGCTGCGCAGGCTGCAGCTCCTGCCGAGTCTGTGTTCGTTCCCTTTTGTCCGGA
GCTCGGAAGAGAGAAGGACAATGAAGCGACGTATCGACCCATCTTCATTTCCAAGACCTT
CTCAGACAACGGGGTTCCCTACGACTTTGTGGTTCTCGAGAAGAGAAGGAAGACTGACGA
CGCAGCCACTGCGGAACCGAGCAACGCAATGAGCTCCTTGACGTCCACGAGGGAGACAAC
TCCCGTGCACGGGTTGCAGGCTCCTTCTTCGGCCGCAGCCATTGCCCCGGTGTTGGCGTG
GATGGACGAAGAAGACCGGAAAAAACGCGAGCAAAAGGAACTGATTCGGGCCGTTCCGCA
TGTTCACTTTAGAGGCCATGAAGAGTTCCAGTACCTTGATCTCATTGCCGACATTATTAA
CAATGGAAGGACAATGGATGACCGAACGGGCGTTGGTGTCATCTCCAAATTCGGCTGCAC
TATGCGCTACTCGCTGGATCAGGCCTTTCCACTTCTCACCACAAAGCGTGTGTTCTGGAA
AGGGGTCCTCGAAGAGTTGCTGTGGTTCATTCGCGGCGACACGAACGCAAACCATCTTTC
TGAGAAGGGCGTGAAGATCTGGGACAAGAATGTGACACGCGAGTTCCTCGATTCGCGCAA
TCTCCCCCACCGAGAGGTCGGAGACATCGGCCCGGGCTACGGCTTCCAGTGGAGACACTT
CGGCGCGGCATACAAAGACATGCACACAGACTACACAGGGCAGGGCGTCGACCAGCTGAA
GAATGTGATCCAGATGCTGAGAACGAATCCAACAGATCGTCGCATGCTCATGACTGCCTG
GAATCCTGCAGCGCTGGACGAAATGGCGCTGCCGCCTTGTCACTTGTTGTGCCAGTTCTA
CGTGAACGACCAGAAGGAGCTGTCGTGCATCATGTATCAGCGGTCGTGCGATGTCGGCCT
CGGCGTCCCCTTCAACATCGCTTCCTATTCGCTTTTGACGCTCATGGTTGCACACGTCTG
CAACCTAAAACCTAAGGAGTTCATTCACTTCATGGGGAACACGCATGTCTACACGAACCA
TGTCGAGGCTTTAAAAGAGCAGCTGCGGAGAGAACCGAGACCGTTCCCCATTGTGAACAT
CCTCAACAAGGAACGCATCAAGGAAATCGACGATTTCACCGCCGAGGATTTTGAGGTCGT
GGGCTACGTCCCGCACGGACGAATCCAGATGGAGATGGCTGTCTAGCGGAAATACAGAAG
CTAGCTTTGATCCCGTTTTTCTTACTTATATATTTATACCAATTGATTGTATTTATAACT
GTAAAAATGTGTATGTTGTGTGCATATTTTTTTTTGTGCATGCACATGCATGTAAATAGC
TAAAATTATGAACATTTTATTTTTTGTTCAGAAAAAAAAAACTTTACACACATAAAATGG
CTAGTATGAATAGCCATATTTTATATAAATTAAATCCTATGAATTTATGACCATATTAAA
AATTTAGATATTTATGGAACATAATATGTTTGAAACAATAAGACAAAATTATTATTATTA
TTATTATTTTTACTGTTATAATTATGTTGTCTCTTCAATGATTCATAAATAGTTGGACTT
GATTTTTAAAATGTTTATAATATGATTAGCATAGTTAAATAAAAAAAGTTGAAAAATTAA
AAAAAAACATATAAACACAAATGATGTTTTTTCCTTCAATTTCGATATCGAATTCAGAAA
ATGCATTTTGAAGATTTAAAAAATATTTTTTGTCAAAATAAAGAAGATTTATGTACAAAT
TTTTATAAACTTATTTATTTTTGGATGAACATGAAAGACGATGAAATATATGATAGCCAA
TTTGTTCGAATTAAGACAGATTTTCAGAATAATATGAATAAATTATTACCTCAATTTTAT
ACAATAACTCCACATTTGTTCTATGAAAATTTTATAGTGGTATAGAAAATAACAAAATAT
AATAGTAATGTGTGTCTATAAGTGTTTAACTTTTCGGTTTGATATTATCATTTATTTTAT
TTTCATGTATATGCAGATGGACCCGTCTCATAACATAACGAGCAGTAGGATCGGATTTTT
TCTTCATAATTATCCAACAAATCAAGATAAAAACATACAATTATTAAAAAAAATAGAAAA
AATTGTAAAAAATAGCAATATCAAAGATGTATATTTTTATTCAGATACATATGTATTGTA
TAATCGCGGCCGCCACCGCGGTGGAGCTCCAATTCGCCCTATAGTGAGTCGTATTACGCG
CGCTCACTGGCCGTCGTTTTACAACGTCGTGACTGGGAAAACCCTGGCGTTACCCAACTT
AATCGCCTTGCAGCACATCCCCCTTTCGCCAGCTGGCGTAATAGCGAAGAGGCCCGCACC
GATCGCCCTTCCCAACAGTTGCGCAGCCTGAATGGCGAATGGGACGCGCCCTGTAGCGGC
GCATTAAGCGCGGCGGGTGTGGTGGTTACGCGCAGCGTGACCGCTACACTTGCCAGCGCC
CTAGCGCCCGCTCCTTTCGCTTTCTTCCCTTCCTTTCTCGCCACGTTCGCCGGCTTTCCC
CGTCAAGCTCTAAATCGGGGGCTCCCTTTAGGGTTCCGATTTAGTGCTTTACGGCACCTC
GACCCCAAAAAACTTGATTAGGGTGATGGTTCACGTAGTGGGCCATCGCCCTGATAGACG
GTTTTTCGCCCTTTGACGTTGGAGTCCACGTTCTTTAATAGTGGACTCTTGTTCCAAACT
GGAACAACACTCAACCCTATCTCGGTCTATTCTTTTGATTTATAAGGGATTTTGCCGATT
TCGGCCTATTGGTTAAAAAATGAGCTGATTTAACAAAAATTTAACGCGAATTTTAACAAA
ATATTAACGCTTACAATTTAGGTGGCACTTTTCGGGGAAATGTGCGCGGAACCCCTATTT
GTTTATTTTTCTAAATACATTCAAATATGTATCCGCTCATGAGACAATAACCCTGATAAA
TGCTTCAATAATATTGAAAAAGGAAGAGTATGAGTATTCAACATTTCCGTGTCGCCCTTA
TTCCCTTTTTTGCGGCATTTTGCCTTCCTGTTTTTGCTCACCCAGAAACGCTGGTGAAAG
TAAAAGATGCTGAAGATCAGTTGGGTGCACGAGTGGGTTACATCGAACTGGATCTCAACA
GCGGTAAGATCCTTGAGAGTTTTCGCCCCGAAGAACGTTTTCCAATGATGAGCACTTTTA
AAGTTCTGCTATGTGGCGCGGTATTATCCCGTATTGACGCCGGGCAAGAGCAACTCGGTC
GCCGCATACACTATTCTCAGAATGACTTGGTTGAGTACTCACCAGTCACAGAAAAGCATC
TTACGGATGGCATGACAGTAAGAGAATTATGCAGTGCTGCCATAACCATGAGTGATAACA
CTGCGGCCAACTTACTTCTGACAACGATCGGAGGACCGAAGGAGCTAACCGCTTTTTTGC
ACAACATGGGGGATCATGTAACTCGCCTTGATCGTTGGGAACCGGAGCTGAATGAAGCCA
TACCAAACGACGAGCGTGACACCACGATGCCTGTAGCAATGGCAACAACGTTGCGCAAAC
TATTAACTGGCGAACTACTTACTCTAGCTTCCCGGCAACAATTAATAGACTGGATGGAGG
CGGATAAAGTTGCAGGACCACTTCTGCGCTCGGCCCTTCCGGCTGGCTGGTTTATTGCTG
ATAAATCTGGAGCCGGTGAGCGTGGGTCTCGCGGTATCATTGCAGCACTGGGGCCAGATG
GTAAGCCCTCCCGTATCGTAGTTATCTACACGACGGGGAGTCAGGCAACTATGGATGAAC
GAAATAGACAGATCGCTGAGATAGGTGCCTCACTGATTAAGCATTGGTAACTGTCAGACC
AAGTTTACTCATATATACTTTAGATTGATTTAAAACTTCATTTTTAATTTAAAAGGATCT
AGGTGAAGATCCTTTTTGATAATCTCATGACCAAAATCCCTTAACGTGAGTTTTCGTTCC
ACTGAGCGTCAGACCCCGTAGAAAAGATCAAAGGATCTTCTTGAGATCCTTTTTTTCTGC
GCGTAATCTGCTGCTTGCAAACAAAAAAACCACCGCTACCAGCGGTGGTTTGTTTGCCGG
ATCAAGAGCTACCAACTCTTTTTCCGAAGGTAACTGGCTTCAGCAGAGCGCAGATACCAA
ATACTGTCCTTCTAGTGTAGCCGTAGTTAGGCCACCACTTCAAGAACTCTGTAGCACCGC
CTACATACCTCGCTCTGCTAATCCTGTTACCAGTGGCTGCTGCCAGTGGCGATAAGTCGT
GTCTTACCGGGTTGGACTCAAGACGATAGTTACCGGATAAGGCGCAGCGGTCGGGCTGAA
CGGGGGGTTCGTGCACACAGCCCAGCTTGGAGCGAACGACCTACACCGAACTGAGATACC
TACAGCGTGAGCTATGAGAAAGCGCCACGCTTCCCGAAGGGAGAAAGGCGGACAGGTATC
CGGTAAGCGGCAGGGTCGGAACAGGAGAGCGCACGAGGGAGCTTCCAGGGGGAAACGCCT
GGTATCTTTATAGTCCTGTCGGGTTTCGCCACCTCTGACTTGAGCGTCGATTTTTGTGAT
GCTCGTCAGGGGGGCGGAGCCTATGGAAAAACGCCAGCAACGCGGCCTTTTTACGGTTCC
TGGCCTTTTGCTGGCCTTTTGCTCACATGTTCTTTCCTGCGTTATCCCCTGATTCTGTGG
ATAACCGTATTACCGCCTTTGAGTGAGCTGATACCGCTCGCCGCAGCCGAACGACCGAGC
GCAGCGAGTCAGTGAGCGAGGAAGCGGAAGAGCGCCCAATACGCAAACCGCCTCTCCCCG
CGCGTTGGCCGATTCATTAATGCAGCTGGCACGACAGGTTTCCCGACTGGAAAGCGGGCA
GTGAGCGCAACGCAATTAATGTGAGTTAGCTCACTCATTAGGCACCCCAGGCTTTACACT
TTATGCTTCCGGCTCGTATGTTGTGTGGAATTGTGA
Restriction sites to linearize plasmid
Partial or complete disruption of the genePartial
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 genetic modification results in the disruption of the following gene models PB101363.00.0 and PB000829.02.0 and PB105739.00.0 (= PF14_0435).
See for more information about the disruption Lasonder, E. et al., 2008, PloS Pathogens 10, e10000195 (Supporting Information: S3).
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 1AAAGGTACCGGTTTAATGTTGGTAGAC
Additional information primer 1L2666 (Asp718); 5'
Sequence Primer 2AAAAAGCTTGAGAATGGTTATAAGAC
Additional information primer 2L2653 (HindIII); 5'
Sequence Primer 3AAAGAATTCAGAAAATGCATTTTGAAG
Additional information primer 3L2654 (EcoRI); 3'
Sequence Primer 4AAAGCGGCCGCGATTATACAATACATATG
Additional information primer 4L2655 (NotI); 3'
Sequence Primer 5
Additional information primer 5
Sequence Primer 6
Additional information primer 6