RMgmDB - Rodent Malaria genetically modified Parasites

Summary

RMgm-502
Malaria parasiteP. berghei
Genotype
DisruptedGene model (rodent): PBANKA_0701100; Gene model (P.falciparum): Not available; Gene product: conserved Plasmodium protein, unknown function
PhenotypeNo phenotype has been described
Last modified: 21 December 2011, 09:53
  *RMgm-502
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) : 22184632
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 1037m1f1mocl1 (RMgm-32)
Other information parent lineP. berghei ANKA 1037m1f1mocl1 (1037cl1; RMgm-32) is a reference ANKA mutant line which expresses GFP-luciferase under control of a schizont-specific promoter (PubMed: PMID: 20019192). It does not contain a drug-selectable marker and has been selected by FACS sorting based on GFP expression
The mutant parasite was generated by
Name PI/ResearcherJ. Fonager, E.M. Pasini, C.J. Janse, B.M Franke-Fayard
Name Group/DepartmentLeiden Malaria Research Group
Name InstituteLeiden University Medical Center (LUMC)
CityLeiden
CountryThe Netherlands
Name of the mutant parasite
RMgm numberRMgm-502
Principal nameexp. 1158
Alternative name
Standardized name
Is the mutant parasite cloned after genetic modificationYes
Phenotype
Asexual blood stageNot different from wild type
Gametocyte/GameteNot tested
Fertilization and ookineteNot tested
OocystNot tested
SporozoiteNot tested
Liver stageNot tested
Additional remarks phenotype

A pyrimethamine-resistant parasite population was selected after transfection with a construct for targeted gene disruption by double cross-over homologous recombination. The primers of the two target sequences of the gene are provided below.

The genotype of the parasites has not been analysed in detail.
Diagnostic PCR (for correct integration of the construct) and/or Southern analysis of PFG-separated chromosomes (to show integration into the chromosome on which the target gene is located) have been performed (see below). This analysis provided evidence for correct disruption of the gene indicating that it has no essential function during asexual blood stage development. However, no further analyses on the parasite population/cloned lines have been performed to show absence of transcripts or protein .

The phenotype of the selected parasite population/cloned lines has not been analysed in detail.
No obvious growth defects of the asexual blood stages of the selected parasite populations/cloned lines have been observed.

The gene has been selected for targeted gene disruption based on its presence in a proteome of membranes of infected red blood cells


  Disrupted: Mutant parasite with a disrupted gene
Details of the target gene
Gene Model of Rodent Parasite PBANKA_0701100
Gene Model P. falciparum ortholog Not available
Gene productconserved Plasmodium 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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GGGCCCCCCCTCGAGGTCGACGGTATCGATAAGCTTGCATGCCTGCAGGTCAACAATAAA
TAATAAATAAATATTGTGGAAATAAAATAACATATAATTATTTTTAATACATTGATTTCC
CTTTTATTTTTTTAAATTTCATTGATATAAAAATATATAATAATAACATATATGATTTCA
AATTAATCTTTTCAAAAATGGTGTTTATTTTTGTATGTTTGTGTATGAATTAATCACATA
ACACATCTATTAAATTGAGTTGGTAATATAGACACAAATAAATATATATATTTTTATAGC
TTAAAAGTGTGTTATGAATATTTTAAGCATATTTTCTTTTTCTTTGGATTGTGTAAAATG
AACTCATATAATGCGTTTTTTTGTTTTTGTTATTTTGTCATTTTGTTATTTTGCTATTTT
ATGGATTAATTTTTGTTTATAAAATGGGAAATAATTTTAACATATTTAAATAAATGGAGA
AAAAATATAAAATAATTATAAAAAAAAGTTAATACACATTTTTTCCTGTTATAGACCTTA
TATTTATTTATCCATATATATATATATATATATATATATATATATATACATACCAAGTGA
ATTAAGAGGAAAGCTAATTTATTATTCAGAATAATATATGAACTATATATAATTTTTATT
ATTTTGGTGTATATTAATCTGTCTATATGCATACATGCAATAATTTATCGACTTATATAT
CAAATAACATAAAATAGAAGTGTTTTAAATTATGGATATATGCTCAATATTCATTTTTTT
TAATAAGTTAGCTATATTTAAATTATACATTTTATATATGGTCTCTTTTTTTTTTAAATA
TTATTTAAGTGATCATGAAAATATAAATAATTTTTTTTTATTTAATATCCTTTTGCTTGC
ATGTGGTAAATGGAAATTTGGATGTGTTTTGAARGTTCGGATATAGTTGTATGGACATAT
AATATATTTTGTGAAAAATTGGTTTTATGTTTATACTTATGCCAATACTTTTTGAGTAAA
ACAAAGCAAGTGCTTATAAATAATTAAAGCCAATTTTATAATATATATTTTTTTATTTAA
TTTGAATTTAGTAGTATAATTTTTTATGGTAAGTGCTCAAAGAGAGTTGCTTATAAAGTA
TGGTTTGTTTCTTTTTCGCCATTTTGAATTACACATTAAAAATATATAGATACATATATT
ATAATATGAAATCATTAATAATTTAGGGAAATTCTACAAATTTAAAAACGAATAAAATAA
TTGTTTTTCATCATGCCATAACACAATATTGATATATACATGTACAAACATTTTTTTTAT
TTGGAAAATATAAATTATATAAAAAAAAATGTATAGTATACAAAATGAGCATATTCACAC
GGGGTGGACGTTCATTTTTTCATTTTTCCCCTGTTTTTTATGAGTATATGATAAAATTTT
ATGAACATTTACACAAAATGAAAATGGATATATAGGAAAAATGGAGCGGTATTTCATTTA
TCTTTGATTGTCATTTGGATATTATATTACCYTGGGTAGGCAATTAAAAATGTTAAATAA
CAATTTAAGGAAATTATATTTTATATATTAAAATTAACACTGTATTATATGATTCGCTTA
TAAAAGCCACTCTTTCCCCATGCAAAGCTGTTTAATATCAATTTTAACAAATTACACACA
TGTTAATATATTTATATATATAATTTATATATTTATAATTTATATATTTATATTTTTATT
ATTTATATATTTATTATTTATTGTGTGTGTCAATTCGGGTAGGATATACCTCTTTTTTAT
TGTTTAAAGCGATTTGTATTCTAAAATATAAAGRATTTGAAAAAGAGAAAGATAGAATAT
GATCCCATCATATATAGCCCTATAATTTTTATTTAGCAGCGAATTAATTTTTCTATTAAG
TTTATGTGTAATTAAAATAACGGAATATATATAATACAATAAAAAAGTGCATAAATTAAA
ATTTTTTCAATTAAATTTTTTTTTTTAAGGGGTTATATAATATTAAATATATAAAATACG
ATTATATATTTTTGCTACAATTTTTTATATTAAGATATAAATAGTAAATAAATGGTATTA
TATGGCATGTAATATATAAATTTTTTCCAATTTTTATTTTATATACACTTTTCCTTTTTT
TGTCATAAAACTTAAACAATTTACACATTCATTTTAAAAATTGACTATTTGTTTCAACAT
TTTTTGAGTTTCCGTTTTATAATAGTATTTTCATTTGTATATTGCTTATATATATAAATA
CACACCTAAATGTTACAAAGGATCAATGCATAAACCGGTGTGTCTGGTCGTCGCGATGAC
CCCCAAGAGGGGCATCGGCATCAACAACGGCCTCCCGTGGCCCCACTTGACCACAGATTT
CAAACACTTTTCTCGTGTGACAAAAACGACGCCCGAAGAAGCCAGTCGCCTGAACGGGTG
GCTTCCCAGGAAATTTGCAAAGACGGGCGACTCTGGACTTCCCTCTCCATCAGTCGGCAA
GAGATTCAACGCCGTTGTCATGGGACGGAAAACCTGGGAAAGCATGCCTCGAAAGTTTAG
ACCCCTCGTGGACAGATTGAACATCGTCGTTTCCTCTTCCCTCAAAGAAGAAGACATTGC
GGCGGAGAAGCCTCAAGCTGAAGGCCAGCAGCGCGTCCGAGTCTGTGCTTCACTCCCAGC
AGCTCTCAGCCTTCTGGAGGAAGAGTACAAGGATTCTGTCGACCAGATTTTTGTCGTGGG
AGGAGCGGGACTGTACGAGGCAGCGCTGTCTCTGGGCGTTGCCTCTCACCTGTACATCAC
GCGTGTAGCCCGCGAGTTTCCGTGCGACGTTTTCTTCCCTGCGTTCCCCGGAGATGACAT
TCTTTCAAACAAATCAACTGCTGCGCAGGCTGCAGCTCCTGCCGAGTCTGTGTTCGTTCC
CTTTTGTCCGGAGCTCGGAAGAGAGAAGGACAATGAAGCGACGTATCGACCCATCTTCAT
TTCCAAGACCTTCTCAGACAACGGGGTTCCCTACGACTTTGTGGTTCTCGAGAAGAGAAG
GAAGACTGACGACGCAGCCACTGCGGAACCGAGCAACGCAATGAGCTCCTTGACGTCCAC
GAGGGAGACAACTCCCGTGCACGGGTTGCAGGCTCCTTCTTCGGCCGCAGCCATTGCCCC
GGTGTTGGCGTGGATGGACGAAGAAGACCGGAAAAAACGCGAGCAAAAGGAACTGATTCG
GGCCGTTCCGCATGTTCACTTTAGAGGCCATGAAGAGTTCCAGTACCTTGATCTCATTGC
CGACATTATTAACAATGGAAGGACAATGGATGACCGAACGGGCGTTGGTGTCATCTCCAA
ATTCGGCTGCACTATGCGCTACTCGCTGGATCAGGCCTTTCCACTTCTCACCACAAAGCG
TGTGTTCTGGAAAGGGGTCCTCGAAGAGTTGCTGTGGTTCATTCGCGGCGACACGAACGC
AAACCATCTTTCTGAGAAGGGCGTGAAGATCTGGGACAAGAATGTGACACGCGAGTTCCT
CGATTCGCGCAATCTCCCCCACCGAGAGGTCGGAGACATCGGCCCGGGCTACGGCTTCCA
GTGGAGACACTTCGGCGCGGCATACAAAGACATGCACACAGACTACACAGGGCAGGGCGT
CGACCAGCTGAAGAATGTGATCCAGATGCTGAGAACGAATCCAACAGATCGTCGCATGCT
CATGACTGCCTGGAATCCTGCAGCGCTGGACGAAATGGCGCTGCCGCCTTGTCACTTGTT
GTGCCAGTTCTACGTGAACGACCAGAAGGAGCTGTCGTGCATCATGTATCAGCGGTCGTG
CGATGTCGGCCTCGGCGTCCCCTTCAACATCGCTTCCTATTCGCTTTTGACGCTCATGGT
TGCACACGTCTGCAACCTAAAACCTAAGGAGTTCATTCACTTCATGGGGAACACGCATGT
CTACACGAACCATGTCGAGGCTTTAAAAGAGCAGCTGCGGAGAGAACCGAGACCGTTCCC
CATTGTGAACATCCTCAACAAGGAACGCATCAAGGAAATCGACGATTTCACCGCCGAGGA
TTTTGAGGTCGTGGGCTACGTCCCGCACGGACGAATCCAGATGGAGATGGCTGTCTAGCG
GAAATACAGAAGCTAGCTTTGATCCCGTTTTTCTTACTTATATATTTATACCAATTGATT
GTATTTATAACTGTAAAAATGTGTATGTTGTGTGCATATTTTTTTTTGTGCATGCACATG
CATGTAAATAGCTAAAATTATGAACATTTTATTTTTTGTTCAGAAAAAAAAAACTTTACA
CACATAAAATGGCTAGTATGAATAGCCATATTTTATATAAATTAAATCCTATGAATTTAT
GACCATATTAAAAATTTAGATATTTATGGAACATAATATGTTTGAAACAATAAGACAAAA
TTATTATTATTATTATTATTTTTACTGTTATAATTATGTTGTCTCTTCAATGATTCATAA
ATAGTTGGACTTGATTTTTAAAATGTTTATAATATGATTAGCATAGTTAAATAAAAAAAG
TTGAAAAATTAAAAAAAAACATATAAACACAAATGATGTTTTTTCCTTCAATTTCGATAT
CGAATTCCATCATAATACTAAAAATGCACTTAACCGTTTGATTACGAACAAATGGAAATA
AGAATATTTATTTAAAATATTAAAAAATAAAATTCTATGTTTTTAACCCCAATATATACA
TGAATATAACAGCCATATTTTTATTATACCGATGATTTTACAGAATAAAGAAATTCAAAA
ATATATTAACAAAAAATGGGTGTCTTCATTAAATTAAAATTTTTTTTTAATAATGGAAAA
GTACCTTCATTTTTAAAATATATTTTTATATTTAGTATTTCTTTTATTGTAATTTCATAT
CAGAATACGCCTTGGTATATTATATGTAAGAATTTTGTTTATACATATATGCGCACATGC
GCTTATTTGTATTTACTCAATTCTTGTGAAACAATTAAGTATTCCTATAACTTTATTGAA
TAATTAATTGTTTTTTATAAAATATGGAATTTCCACTTACCATGAAATGACATTTTTAAG
AAGCAAAAAGTTTACATATATAATTAGAAAAAAATAGATACATTCACATGCATATTATAT
ATATTTGATACAATGTGAATTATACCGTAGATGTTTTTTTTATGCCGGATCCACTAGTTC
TAGAGCGGCCGCCACCGCGGTGGAGCTCCAATTCGCCCTATAGTGAGTCGTATTACAATT
CACTGGCCGTCGTTTTACAACGTCGTGACTGGGAAAACCCTGGCGTTACCCAACTTAATC
GCCTTGCAGCACATCCCCCTTTCGCCAGCTGGCGTAATAGCGAAGAGGCCCGCACCGATC
GCCCTTCCCAACAGTTGCGCAGCCTGAATGGCGAATGGGACGCGCCCTGTAGCGGCGCAT
TAAGCGCGGCGGGTGTGGTGGTTACGCGCAGCGTGACCGCTACACTTGCCAGCGCCCTAG
CGCCCGCTCCTTTCGCTTTCTTCCCTTCCTTTCTCGCCACGTTCGCCGGCTTTCCCCGTC
AAGCTCTAAATCGGGGGCTCCCTTTAGGGTTCCGATTTAGTGCTTTACGGCACCTCGACC
CCAAAAAACTTGATTAGGGTGATGGTTCACGTAGTGGGCCATCGCCCTGATAGACGGTTT
TTCGCCCTTTGACGTTGGAGTCCACGTTCTTTAATAGTGGACTCTTGTTCCAAACTGGAA
CAACACTCAACCCTATCTCGGTCTATTCTTTTGATTTATAAGGGATTTTGCCGATTTCGG
CCTATTGGTTAAAAAATGAGCTGATTTAACAAAAATTTAACGCGAATTTTAACAAAATAT
TAACGCTTACAATTTAGGTGGCACTTTTCGGGGAAATGTGCGCGGAACCCCTATTTGTTT
ATTTTTCTAAATACATTCAAATATGTATCCGCTCATGAGACAATAACCCTGATAAATGCT
TCAATAATATTGAAAAAGGAAGAGTATGAGTATTCAACATTTCCGTGTCGCCCTTATTCC
CTTTTTTGCGGCATTTTGCCTTCCTGTTTTTGCTCACCCAGAAACGCTGGTGAAAGTAAA
AGATGCTGAAGATCAGTTGGGTGCACGAGTGGGTTACATCGAACTGGATCTCAACAGCGG
TAAGATCCTTGAGAGTTTTCGCCCCGAAGAACGTTTTCCAATGATGAGCACTTTTAAAGT
TCTGCTATGTGGCGCGGTATTATCCCGTATTGACGCCGGGCAAGAGCAACTCGGTCGCCG
CATACACTATTCTCAGAATGACTTGGTTGAGTACTCACCAGTCACAGAAAAGCATCTTAC
GGATGGCATGACAGTAAGAGAATTATGCAGTGCTGCCATAACCATGAGTGATAACACTGC
GGCCAACTTACTTCTGACAACGATCGGAGGACCGAAGGAGCTAACCGCTTTTTTGCACAA
CATGGGGGATCATGTAACTCGCCTTGATCGTTGGGAACCGGAGCTGAATGAAGCCATACC
AAACGACGAGCGTGACACCACGATGCCTGTAGCAATGGCAACAACGTTGCGCAAACTATT
AACTGGCGAACTACTTACTCTAGCTTCCCGGCAACAATTAATAGACTGGATGGAGGCGGA
TAAAGTTGCAGGACCACTTCTGCGCTCGGCCCTTCCGGCTGGCTGGTTTATTGCTGATAA
ATCTGGAGCCGGTGAGCGTGGGTCTCGCGGTATCATTGCAGCACTGGGGCCAGATGGTAA
GCCCTCCCGTATCGTAGTTATCTACACGACGGGGAGTCAGGCAACTATGGATGAACGAAA
TAGACAGATCGCTGAGATAGGTGCCTCACTGATTAAGCATTGGTAACTGTCAGACCAAGT
TTACTCATATATACTTTAGATTGATTTAAAACTTCATTTTTAATTTAAAAGGATCTAGGT
GAAGATCCTTTTTGATAATCTCATGACCAAAATCCCTTAACGTGAGTTTTCGTTCCACTG
AGCGTCAGACCCCGTAGAAAAGATCAAAGGATCTTCTTGAGATCCTTTTTTTCTGCGCGT
AATCTGCTGCTTGCAAACAAAAAAACCACCGCTACCAGCGGTGGTTTGTTTGCCGGATCA
AGAGCTACCAACTCTTTTTCCGAAGGTAACTGGCTTCAGCAGAGCGCAGATACCAAATAC
TGTCCTTCTAGTGTAGCCGTAGTTAGGCCACCACTTCAAGAACTCTGTAGCACCGCCTAC
ATACCTCGCTCTGCTAATCCTGTTACCAGTGGCTGCTGCCAGTGGCGATAAGTCGTGTCT
TACCGGGTTGGACTCAAGACGATAGTTACCGGATAAGGCGCAGCGGTCGGGCTGAACGGG
GGGTTCGTGCACACAGCCCAGCTTGGAGCGAACGACCTACACCGAACTGAGATACCTACA
GCGTGAGCTATGAGAAAGCGCCACGCTTCCCGAAGGGAGAAAGGCGGACAGGTATCCGGT
AAGCGGCAGGGTCGGAACAGGAGAGCGCACGAGGGAGCTTCCAGGGGGAAACGCCTGGTA
TCTTTATAGTCCTGTCGGGTTTCGCCACCTCTGACTTGAGCGTCGATTTTTGTGATGCTC
GTCAGGGGGGCGGAGCCTATGGAAAAACGCCAGCAACGCGGCCTTTTTACGGTTCCTGGC
CTTTTGCTGGCCTTTTGCTCACATGTTCTTTCCTGCGTTATCCCCTGATTCTGTGGATAA
CCGTATTACCGCCTTTGAGTGAGCTGATACCGCTCGCCGCAGCCGAACGACCGAGCGCAG
CGAGTCAGTGAGCGAGGAAGCGGAAGAGCGCCCAATACGCAAACCGCCTCTCCCCGCGCG
TTGGCCGATTCATTAATGCAGCTGGCACGACAGGTTTCCCGACTGGAAAGCGGGCAGTGA
GCGCAACGCAATTAATGTGAGTTAGCTCACTCATTAGGCACCCCAGGCTTTACACTTTAT
GCTTCCGGCTCGTATGTTGTGTGGAATTGTGAGCGGATAACAATTTCACACAGGAAACAG
CTATGACCATGATTACGCCAAGCTCGAAATTAACCCTCACTAAAGGGAACAAAAGCTGGT
ACCTCTAGAGGTACAAAAATGGAAATGTATATAGCCTAAATTATTTTTTAAACGGCTTAA
TATAATATGCAATAGTAACACAATTATTATATATGTTTTATATATATGAAAAAAATGCAT
ACGTCATCTAAATATTTATTTTTCTTCATCAATAAAATATATCCTAAAATATTAGGGTAC
AAAAGAAAATTTTTATTTTAATATATTATTATTTTATCCCACTTTCGTATTTACTTATTA
TTAATATATAATAATTACAAATAATGGTATTTTTATCATCAAGATTTATGTTTATTTTTA
TTATGCACAAAAAATAATATATAATAAAATTGTATAAGATACATATTTACATGAGTAGTA
TTAGTTTTACTATTTTACTATAGTTTTTATTCATTTATAATTTTTTTATATTGTAAAATA
TTATTATTACTATTATTATTTTATTAGAGTTTTGTAAATATTTATTTAAATTCCTAATAT
TTTTATTTACATTATATATAGCTAAAAATAATATTAATGAATTTTTAAGCTTCAAAAGGC
ATTTAATATTTATCCTATTATTTTGTTATATTAATATTTAACTTTTTTAATTATTATATT
CTACAATATGAAGCACAAATTGCATACCTTTTTTGTTATCATATGCTTAGTCGTAGTATT
GTATTGTGGTGGTATAGAGGGAAAACAAAATAATAATATAAAAAAGGATATGTGGATATC
CCGATTTTTTACTCGGTACC
Restriction sites to linearize plasmid Asp 718I/XbaI/BamHI
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 modification
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 1GCCGGTACCTCTAGAGGTACAAAAATGGAAATGTATATAGCC
Additional information primer 1F1 primer
Sequence Primer 2GCCGGTACCGAGTAAAAAATCGGGATATCCAC
Additional information primer 2R1 primer
Sequence Primer 3GCCGAATTCCATCATAATACTAAAAATGCAC
Additional information primer 3F2 primer
Sequence Primer 4GCCGGATCCGGCATAAAAAAAACATCTACG
Additional information primer 4R2 primer
Sequence Primer 5
Additional information primer 5
Sequence Primer 6
Additional information primer 6