RMgmDB - Rodent Malaria genetically modified Parasites

Back to search results

Summary

RMgm-713
Malaria parasiteP. berghei
Genotype
TaggedGene model (rodent): PBANKA_0623100; Gene model (P.falciparum): Not available; Gene product: Plasmodium exported protein, unknown function
Name tag: mCherry
Transgene
Transgene not Plasmodium: A fusion of GFP (gfp-mu3) and Luciferase Firefly (LucIAV)
Promoter: Gene model: PBANKA_0915000; Gene model (P.falciparum): PF3D7_1133400; Gene product: apical membrane antigen 1 (AMA-1)
3'UTR: Gene model: PBANKA_0719300; Gene product: bifunctional dihydrofolate reductase-thymidylate synthase, putative (dhfr/ts)
Replacement locus: Gene model: PBANKA_0306000; Gene product: 6-cysteine protein (230p)
Phenotype Asexual bloodstage;
Last modified: 2 December 2012, 15:41
  *RMgm-713
Successful modificationThe parasite was generated by the genetic modification
The mutant contains the following genetic modification(s) Gene tagging, Introduction of a transgene
Reference (PubMed-PMID number) Reference 1 (PMID number) : 23197789
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. This reference line does not contain a drug-selectable marker (PubMed: PMID: 20019192).
The mutant parasite was generated by
Name PI/ResearcherE.M. Pasini, C.J. Janse, B.M.D. Franke-Fayard
Name Group/DepartmentDepartment of Parasitology
Name InstituteBiomedical Primate Research Centre
CityRijswijk
CountryThe Netherlands
Name of the mutant parasite
RMgm numberRMgm-713
Principal name1622
Alternative name
Standardized name
Is the mutant parasite cloned after genetic modificationNo
Phenotype
Asexual blood stageThe tagged-protein is exported into the cytoplasm of the host erythrocyte and shows a punctuate, vesicle-like localization in the cytoplasm
Gametocyte/GameteNot tested
Fertilization and ookineteNot tested
OocystNot tested
SporozoiteNot tested
Liver stageNot tested
Additional remarks phenotype

Mutant/mutation

Protein (function)

Phenotype

Additional information
The mutant parasite expresses an mCherry-tagged protein. The protein has been selected for tagging in a screen for putative exported proteins of P. berghei.

Phenotype analyses of the blood stages by fluorescence microscopy show that the tagged-protein is exported into the cytoplasm of the host erythrocyte and shows a punctuate, vesicle-like localization in the cytoplasm.

Mutant 1622 was generated using a plasmid construct harboring a fragment of PBANKA_062310 , c-terminally fused to mCherry. A second independent mutant (2061) was generated using a plasmid construct in which the entire PBANKA_062310 coding region was c-terminally fused to mCherry. Both constructs aim at integration of the tagging construct by single-cross-over integration resulting in the presence of a tagged copy of the endogenous gene. Southern analysis of PFG-separated chromosomes of both mutants (to show integration into the chromosome on which the target gene is located) unexpectedly shows  integration of the  construct into one of the chromosomes of the group 9/10/11 (see below).



Other mutants
RMgm-504: unsuccessful attempt to disrupt PBANKA_062310


  Tagged: Mutant parasite with a tagged gene
Details of the target gene
Gene Model of Rodent Parasite PBANKA_0623100
Gene Model P. falciparum ortholog Not available
Gene productPlasmodium exported protein, unknown function
Gene product: Alternative name
Details of the genetic modification
Name of the tagmCherry
Details of taggingC-terminal
Additional remarks: tagging
Commercial source of tag-antibodiesClontech
Type of plasmid/constructPlasmid single 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
CTAAATTGTAAGCGTTAATATTTTGTTAAAATTCGCGTTAAATTTTTGTTAAATCAGCTC
ATTTTTTAACCAATAGGCCGAAATCGGCAAAATCCCTTATAAATCAAAAGAATAGACCGA
GATAGGGTTGAGTGTTGTTCCAGTTTGGAACAAGAGTCCACTATTAAAGAACGTGGACTC
CAACGTCAAAGGGCGAAAAACCGTCTATCAGGGCGATGGCCCACTACGTGAACCATCACC
CTAATCAAGTTTTTTGGGGTCGAGGTGCCGTAAAGCACTAAATCGGAACCCTAAAGGGAG
CCCCCGATTTAGAGCTTGACGGGGAAAGCCGGCGAACGTGGCGAGAAAGGAAGGGAAGAA
AGCGAAAGGAGCGGGCGCTAGGGCGCTGGCAAGTGTAGCGGTCACGCTGCGCGTAACCAC
CACACCCGCCGCGCTTAATGCGCCGCTACAGGGCGCGTCCCATTCGCCATTCAGGCTGCG
CAACTGTTGGGAAGGGCGATCGGTGCGGGCCTCTTCGCTATTACGCCAGCTGGCGAAAGG
GGGATGTGCTGCAAGGCGATTAAGTTGGGTAACGCCAGGGTTTTCCCAGTCACGACGTTG
TAAAACGACGGCCAGTGAGCGCGCGTAATACGACTCACTATAGGGCGAATTGGAGCTCCA
CCGCGGTGGCGGCCGCTCTAGAACTAGTAGCAGCATACATGGGATACTTGGTATTGGGGA
GTTCAAAGGTAAATACAATATCATTTAAATTATTTCATTATTCACATATATGCATATACA
TATTATTCATATAAGCTAGTATATATGTTTAAATGCATATGTACTTCTATGTGACGATTA
TATATATTTATTTGCTTATATTCCATATACATATTAATATATTTTATCTTTTTTATAGAA
TAAAAAAAATAAAAAGGGGGATAATAACGTAAATCCAATTCCCAATGTTAAAAATATTTA
CAATAAACGTAAGTATACGGATGATCAATCATCAGCACCAAAAACAGATAAATGGAAACA
TGATGCTTGGAGAAAATGGATTTTAAATTTAGACGATAGATGGAACACATTTAGCAGAAA
TGTTGATAAACATAAAGGTGAATGGATGGACCAAAAAGATGGCCAATTGATAGAATGGGT
AAAAGAATTACAAGCAAAATGGATGCATTTCAATCCGAATATAGATCAAGAATATAAAAT
TGAATTTCTAAAAAAGTCTGAAAACTGGACTGATGATAAATGGAAAATATGGATGTTAAC
AGAAGCACAACAACTTTTAGAAAATGATCTAAAAGAGTGGCTTTCTCAGGCTGAAGCAGA
TTATTGTAAATGGACTATGGATGCATGGAATCAATGGAAAGATAGTCAAATAAAAGAATG
GATAACTCTTGATTGGAAATATGAAGAAGACACTTTCTTTTCGAAAATTGATGAATTTAC
AGTAGAATCATTACCACCAGAAGACCAAAGATTATGGTATTCTTGGAAAGAAAGAATATA
CAAAGAAGGAAGTGATTGGAAATATTGGGCATCACTAAAGGAATCTAAATATGTAAATGA
AAATTGGGAATTATGGGAAGAATGGAAAAATGAAAAACAAGAAATGTTCAGTGAATGGCT
TCAATTATATACTAATAAATGGATCCAACAAAAACAATGGAGATTATGGATTGGAGAAAA
AATGTCACAAAATGATCAAAAACTTACATCTGAATCGATTTCCGTAGGTGCTAGCGCTGG
CGAAAGATCCGGTGGAGGTGGAAGATCCATGGTGAGCAAGGGCGAGGAGGATAACATGGC
CATCATCAAGGAGTTCATGCGCTTCAAGGTGCACATGGAGGGCTCCGTGAACGGCCACGA
GTTCGAGATCGAGGGCGAGGGCGAGGGCCGCCCCTACGAGGGCACCCAGACCGCCAAGCT
GAAGGTGACCAAGGGTGGCCCCCTGCCCTTCGCCTGGGACATCCTGTCCCCTCAGTTCAT
GTACGGCTCCAAGGCCTACGTGAAGCACCCCGCCGACATCCCCGACTACTTGAAGCTGTC
CTTCCCCGAGGGCTTCAAGTGGGAGCGCGTGATGAACTTCGAGGACGGCGGCGTGGTGAC
CGTGACCCAGGACTCCTCCCTGCAGGACGGCGAGTTCATCTACAAGGTGAAGCTGCGCGG
CACCAACTTCCCCTCCGACGGCCCCGTAATGCAGAAGAAGACCATGGGCTGGGAGGCCTC
CTCCGAGCGGATGTACCCCGAGGACGGCGCCCTGAAGGGCGAGATCAAGCAGAGGCTGAA
GCTGAAGGACGGCGGCCACTACGACGCTGAGGTCAAGACCACCTACAAGGCCAAGAAGCC
CGTGCAGCTGCCCGGCGCCTACAACGTCAACATCAAGTTGGACATCACCTCCCACAACGA
GGACTACACCATCGTGGAACAGTACGAACGCGCCGAGGGCCGCCACTCCACCGGCGGCAT
GGACGAGCTGTACAAGTAATCTAGAATTATTAATATATATGAATATATATACATCGTTGT
ATGCGTAATATACGTGTAACTGTTTTTTTTAATTTTTATATTAACTTTTCCTTATACATT
TTAATGCCATTAACTAATAATATTTTACTTGAAAAAATTCTAAATTTTTATAACTCTAAT
ATAAAATCTGTGAATAATAAAAAATTAGCATAAAGTACGAATTAAACATCGAATATTTGT
TTTAAAGGGGCACACATTATGTGCATATAATTTTATTCTTTGAATTTAATTTTTTGCTTA
TAATTAGATTAAATTAATATAATTTTTTTATTGAAAATAAAAACTTATTTTAAAAAATTA
ACAAATGGAAGTTAATGAAAAATACGAAAAAAGTAAAGGGTTAATTCTTATATGGTCGGT
ACCGAGCTCGAATTCGATATCGAAATTGAAGGAAAAAACATCATTTGTGTTTATATGTTT
TTTTTTAATTTTTCAACTTTTTTTATTTAACTATGCTAATCATATTATAAACATTTTAAA
AATCAAGTCCAACTATTTATGAATCATTGAAGAGACAACATAATTATAACAGTAAAAATA
ATAATAATAATAATAATTTTGTCTTATTGTTTCAAACATATTATGTTCCATAAATATCTA
AATTTTTAATATGGTCATAAATTCATAGGATTTAATTTATATAAAATATGGCTATTCATA
CTAGCCATTTTATGTGTGTAAAGTTTTTTTTTTCTGAACAAAAAATAAAATGTTCATAAT
TTTAGCTATTTACATGCATGTGCATGCACAAAAAAAAATATGCACACAACATACACATTT
TTACAGTTATAAATACAATCAATTGGTATAAATATATAAGTAAGAAAAACGGGATCAAAG
CTAGCTTCTGTATTTCCGCTAGACAGCCATCTCCATCTGGATTCGTCCGTGCGGGACGTA
GCCCACGACCTCAAAATCCTCGGCGGTGAAATCGTCGATTTCCTTGATGCGTTCCTTGTT
GAGGATGTTCACAATGGGGAACGGTCTCGGTTCTCTCCGCAGCTGCTCTTTTAAAGCCTC
GACATGGTTCGTGTAGACATGCGTGTTCCCCATGAAGTGAATGAACTCCTTAGGTTTTAG
GTTGCAGACGTGTGCAACCATGAGCGTCAAAAGCGAATAGGAAGCGATGTTGAAGGGGAC
GCCGAGGCCGACATCGCACGACCGCTGATACATGATGCACGACAGCTCCTTCTGGTCGTT
CACGTAGAACTGGCACAACAAGTGACAAGGCGGCAGCGCCATTTCGTCCAGCGCTGCAGG
ATTCCAGGCAGTCATGAGCATGCGACGATCTGTTGGATTCGTTCTCAGCATCTGGATCAC
ATTCTTCAGCTGGTCGACGCCCTGCCCTGTGTAGTCTGTGTGCATGTCTTTGTATGCCGC
GCCGAAGTGTCTCCACTGGAAGCCGTAGCCCGGGCCGATGTCTCCGACCTCTCGGTGGGG
GAGATTGCGCGAATCGAGGAACTCGCGTGTCACATTCTTGTCCCAGATCTTCACGCCCTT
CTCAGAAAGATGGTTTGCGTTCGTGTCGCCGCGAATGAACCACAGCAACTCTTCGAGGAC
CCCTTTCCAGAACACACGCTTTGTGGTGAGAAGTGGAAAGGCCTGATCCAGCGAGTAGCG
CATAGTGCAGCCGAATTTGGAGATGACACCAACGCCCGTTCGGTCATCCATTGTCCTTCC
ATTGTTAATAATGTCGGCAATGAGATCAAGGTACTGGAACTCTTCATGGCCTCTAAAGTG
AACATGCGGAACGGCCCGAATCAGTTCCTTTTGCTCGCGTTTTTTCCGGTCTTCTTCGTC
CATCCACGCCAACACCGGGGCAATGGCTGCGGCCGAAGAAGGAGCCTGCAACCCGTGCAC
GGGAGTTGTCTCCCTCGTGGACGTCAAGGAGCTCATTGCGTTGCTCGGTTCCGCAGTGGC
TGCGTCGTCAGTCTTCCTTCTCTTCTCGAGAACCACAAAGTCGTAGGGAACCCCGTTGTC
TGAGAAGGTCTTGGAAATGAAGATGGGTCGATACGTCGCTTCATTGTCCTTCTCTCTTCC
GAGCTCCGGACAAAAGGGAACGAACACAGACTCGGCAGGAGCTGCAGCCTGCGCAGCAGT
TGATTTGTTTGAAAGAATGTCATCTCCGGGGAACGCAGGGAAGAAAACGTCGCACGGAAA
CTCGCGGGCTACACGCGTGATGTACAGGTGAGAGGCAACGCCCAGAGACAGCGCTGCCTC
GTACAGTCCCGCTCCTCCCACGACAAAAATCTGGTCGACAGAATCCTTGTACTCTTCCTC
CAGAAGGCTGAGAGCTGCTGGGAGTGAAGCACAGACTCGGACGCGCTGCTGGCCTTCAGC
TTGAGGCTTCTCCGCCGCAATGTCTTCTTCTTTGAGGGAAGAGGAAACGACGATGTTCAA
TCTGTCCACGAGGGGTCTAAACTTTCGAGGCATGCTTTCCCAGGTTTTCCGTCCCATGAC
AACGGCGTTGAATCTCTTGCCGACTGATGGAGAGGGAAGTCCAGAGTCGCCCGTCTTTGC
AAATTTCCTGGGAAGCCACCCGTTCAGGCGACTGGCTTCTTCGGGCGTCGTTTTTGTCAC
ACGAGAAAAGTGTTTGAAATCTGTGGTCAAGTGGGGCCACGGGAGGCCGTTGTTGATGCC
GATGCCCCTCTTGGGGGTCATCGCGACGACCAGACACACCGGTTTATGCATTGATCCTTT
GTAACATTTAGGTGTGTATTTATATATATAAGCAATATACAAATGAAAATACTATTATAA
AACGGAAACTCAAAAAATGTTGAAACAAATAGTCAATTTTTAAAATGAATGTGTAAATTG
TTTAAGTTTTATGACAAAAAAAGGAAAAGTGTATATAAAATAAAAATTGGAAAAAATTTA
TATATTACATGCCATATAATACCATTTATTTACTATTTATATCTTAATATAAAAAATTGT
AGCAAAAATATATAATCGTATTTTATATATTTAATATTATATAACCCCTTAAAAAAAAAA
ATTTAATTGAAAAAATTTTAATTTATGCACTTTTTTATTGTATTATATATATTCCGTTAT
TTTAATTACACATAAACTTAATAGAAAAATTAATTCGCTGCTAAATAAAAATTATAGGGC
TATATATGATGGGATCATATTCTATCTTTCTCTTTTTCAAATYCTTTATATTTTAGAATA
CAAATCGCTTTAAACAATAAAAAAGAGGTATATCCTACCCGAATTGACACACACAATAAA
TAATAAATATATAAATAATAAAAATATAAATATATAAATTATAAATATATAAATTATATA
TATAAATATATTAACATGTGTGTAATTTGTTAAAATTGATATTAAACAGCTTTGCATGGG
GAAAGAGTGGCTTTTATAAGCGAATCATATAATACAGTGTTAATTTTAATATATAAAATA
TAATTTCCTTAAATTGTTATTTAACATTTTTAATTGCCTACCCARGGTAATATAATATCC
AAATGACAATCAAAGATAAATGAAATACCGCTCCATTTTTCCTATATATCCATTTTCATT
TTGTGTAAATGTTCATAAAATTTTATCATATACTCATAAAAAACAGGGGAAAAATGAAAA
AATGAACGTCCACCCCGTGTGAATATGCTCATTTTGTATACTATACATTTTTTTTTATAT
AATTTATATTTTCCAAATAAAAAAAATGTTTGTACATGTATATATCAATATTGTGTTATG
GCATGATGAAAAACAATTATTTTATTCGTTTTTAAATTTGTAGAATTTCCCTAAATTATT
AATGATTTCATATTATAATATATGTATCTATATATTTTTAATGTGTAATTCAAAATGGCG
AAAAAGAAACAAACCATACTTTATAAGCAACTCTCTTTGAGCACTTACCATAAAAAATTA
TACTACTAAATTCAAATTAAATAAAAAAATATATATTATAAAATTGGCTTTAATTATTTA
TAAGCACTTGCTTTGTTTTACTCAAAAAGTATTGGCATAAGTATAAACATAAAACCAATT
TTTCACAAAATATATTATATGTCCATACAACTATATCCGAACYTTCAAAACACATCCAAA
TTTCCATTTACCACATGCAAGCAAAAGGATATTAAATAAAAAAAAATTATTTATATTTTC
ATGATCACTTAAATAATATTTAAAAAAAAAAGAGACCATATATAAAATGTATAATTTAAA
TATAGCTAACTTATTAAAAAAAATGAATATTGAGCATATATCCATAATTTAAAACACTTC
TATTTTATGTTATTTGATATATAAGTCGATAAATTATTGCATGTATGCATATAGACAGAT
TAATATACACCAAAATAATAAAAATTATATATAGTTCATATATTATTCTGAATAATAAAT
TAGCTTTCCTCTTAATTCACTTGGTATGTATATATATATATATATATATATATATATATA
TATGGATAAATAAATATAAGGTCTATAACAGGAAAAAATGTGTATTAACTTTTTTTTATA
ATTATTTTATATTTTTTCTCCATTTATTTAAATATGTTAAAATTATTTCCCATTTTATAA
ACAAAAATTAATCCATAAAATAGCAAAATAACAAAATGACAAAATAACAAAAACAAAAAA
ACGCATTATATGAGTTCATTTTACACAATCCAAAGAAAAAGAAAATATGCTTAAAATATT
CATAACACACTTTTAAGCTATAAAAATATATATATTTATTTGTGTCTATATTACCAACTC
AATTTAATAGATGTGTTATGTGATTAATTCATACACAAACATACAAAAATAAACACCATT
TTTGAAAAGATTAATTTGAAATCATATATGTTATTATTATATATTTTTATATCAATGAAA
TTTAAAAAAATAAAAGGGAAATCAATGTATTAAAAATAATTATATGTTATTTTATTTCCA
CAATATTTATTTATTATTTATTGTTGACCTGCAGGCATGCAAGCTTATCGATACCGTCGA
CCTCGAGGGGGGGCCCGGTACCCAGCTTTTGTTCCCTTTAGTGAGGGTTAATTGCGCGCT
TGGCGTAATCATGGTCATAGCTGTTTCCTGTGTGAAATTGTTATCCGCTCACAATTCCAC
ACAACATACGAGCCGGAAGCATAAAGTGTAAAGCCTGGGGTGCCTAATGAGTGAGCTAAC
TCACATTAATTGCGTTGCGCTCACTGCCCGCTTTCCAGTCGGGAAACCTGTCGTGCCAGC
TGCATTAATGAATCGGCCAACGCGCGGGGAGAGGCGGTTTGCGTATTGGGCGCTCTTCCG
CTTCCTCGCTCACTGACTCGCTGCGCTCGGTCGTTCGGCTGCGGCGAGCGGTATCAGCTC
ACTCAAAGGCGGTAATACGGTTATCCACAGAATCAGGGGATAACGCAGGAAAGAACATGT
GAGCAAAAGGCCAGCAAAAGGCCAGGAACCGTAAAAAGGCCGCGTTGCTGGCGTTTTTCC
ATAGGCTCCGCCCCCCTGACGAGCATCACAAAAATCGACGCTCAAGTCAGAGGTGGCGAA
ACCCGACAGGACTATAAAGATACCAGGCGTTTCCCCCTGGAAGCTCCCTCGTGCGCTCTC
CTGTTCCGACCCTGCCGCTTACCGGATACCTGTCCGCCTTTCTCCCTTCGGGAAGCGTGG
CGCTTTCTCATAGCTCACGCTGTAGGTATCTCAGTTCGGTGTAGGTCGTTCGCTCCAAGC
TGGGCTGTGTGCACGAACCCCCCGTTCAGCCCGACCGCTGCGCCTTATCCGGTAACTATC
GTCTTGAGTCCAACCCGGTAAGACACGACTTATCGCCACTGGCAGCAGCCACTGGTAACA
GGATTAGCAGAGCGAGGTATGTAGGCGGTGCTACAGAGTTCTTGAAGTGGTGGCCTAACT
ACGGCTACACTAGAAGGACAGTATTTGGTATCTGCGCTCTGCTGAAGCCAGTTACCTTCG
GAAAAAGAGTTGGTAGCTCTTGATCCGGCAAACAAACCACCGCTGGTAGCGGTGGTTTTT
TTGTTTGCAAGCAGCAGATTACGCGCAGAAAAAAAGGATCTCAAGAAGATCCTTTGATCT
TTTCTACGGGGTCTGACGCTCAGTGGAACGAAAACTCACGTTAAGGGATTTTGGTCATGA
GATTATCAAAAAGGATCTTCACCTAGATCCTTTTAAATTAAAAATGAAGTTTTAAATCAA
TCTAAAGTATATATGAGTAAACTTGGTCTGACAGTTACCAATGCTTAATCAGTGAGGCAC
CTATCTCAGCGATCTGTCTATTTCGTTCATCCATAGTTGCCTGACTCCCCGTCGTGTAGA
TAACTACGATACGGGAGGGCTTACCATCTGGCCCCAGTGCTGCAATGATACCGCGAGACC
CACGCTCACCGGCTCCAGATTTATCAGCAATAAACCAGCCAGCCGGAAGGGCCGAGCGCA
GAAGTGGTCCTGCAACTTTATCCGCCTCCATCCAGTCTATTAATTGTTGCCGGGAAGCTA
GAGTAAGTAGTTCGCCAGTTAATAGTTTGCGCAACGTTGTTGCCATTGCTACAGGCATCG
TGGTGTCACGCTCGTCGTTTGGTATGGCTTCATTCAGCTCCGGTTCCCAACGATCAAGGC
GAGTTACATGATCCCCCATGTTGTGCAAAAAAGCGGTTAGCTCCTTCGGTCCTCCGATCG
TTGTCAGAAGTAAGTTGGCCGCAGTGTTATCACTCATGGTTATGGCAGCACTGCATAATT
CTCTTACTGTCATGCCATCCGTAAGATGCTTTTCTGTGACTGGTGAGTACTCAACCAAGT
CATTCTGAGAATAGTGTATGCGGCGACCGAGTTGCTCTTGCCCGGCGTCAATACGGGATA
ATACCGCGCCACATAGCAGAACTTTAAAAGTGCTCATCATTGGAAAACGTTCTTCGGGGC
GAAAACTCTCAAGGATCTTACCGCTGTTGAGATCCAGTTCGATGTAACCCACTCGTGCAC
CCAACTGATCTTCAGCATCTTTTACTTTCACCAGCGTTTCTGGGTGAGCAAAAACAGGAA
GGCAAAATGCCGCAAAAAAGGGAATAAGGGCGACACGGAAATGTTGAATACTCATACTCT
TCCTTTTTCAATATTATTGAAGCATTTATCAGGGTTATTGTCTCATGAGCGGATACATAT
TTGAATGTATTTAGAAAAATAAACAAATAGGGGTTCCGCGCACATTTCCCCGAAAAGTGC
CAC
Restriction sites to linearize plasmid HpaI
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 1AACTAGTAGCAGCATACATGG
Additional information primer 1L5020 (SpeI); PBANKA_062310 Forward
Sequence Primer 2AAGATCTTTCGCCAGCGCTAG
Additional information primer 2L5019 (BglII); PBANKA_062310 Reverse
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

  Transgene: Mutant parasite expressing a transgene
Type and details of transgene
Is the transgene Plasmodium derived Transgene: not Plasmodium
Transgene nameA fusion of GFP (gfp-mu3) and Luciferase Firefly (LucIAV)
Details of the genetic modification
Inducable system usedNo
Additional remarks inducable system
Type of plasmid/constructPlasmid 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
AATTCACTGGCCGTCGTTTTACAACGTCGTGACTGGGAAAACCCTGGCGTTACCCAACTT
AATCGCCTTGCAGCACATCCCCCTTTCGCCAGCTGGCGTAATAGCGAAGAGGCCCGCACC
GATCGCCCTTCCCAACAGTTGCGCAGCCTGAATGGCGAATGGCGCCTGATGCGGTATTTT
CTCCTTACGCATCTGTGCGGTATTTCACACCGCATATGGTGCACTCTCAGTACAATCTGC
TCTGATGCCGCATAGTTAAGCCAGCCCCGACACCCGCCAACACCCGCTGACGCGCCCTGA
CGGGCTTGTCTGCTCCCGGCATCCGCTTACAGACAAGCTGTGACCGTCTCCGGGAGCTGC
ATGTGTCAGAGGTTTTCACCGTCATCACCGAAACGCGCGAGACGAAAGGGCCTCGTGATA
CGCCTATTTTTATAGGTTAATGTCATGATAATAATGGTTTCTTAGACGTCAGGTGGCACT
TTTCGGGGAAATGTGCGCGGAACCCCTATTTGTTTATTTTTCTAAATACATTCAAATATG
TATCCGCTCATGAGACAATAACCCTGATAAATGCTTCAATAATATTGAAAAAGGAAGAGT
ATGAGTATTCAACATTTCCGTGTCGCCCTTATTCCCTTTTTTGCGGCATTTTGCCTTCCT
GTTTTTGCTCACCCAGAAACGCTGGTGAAAGTAAAAGATGCTGAAGATCAGTTGGGTGCA
CGAGTGGGTTACATCGAACTGGATCTCAACAGCGGTAAGATCCTTGAGAGTTTTCGCCCC
GAAGAACGTTTTCCAATGATGAGCACTTTTAAAGTTCTGCTATGTGGCGCGGTATTATCC
CGTATTGACGCCGGGCAAGAGCAACTCGGTCGCCGCATACACTATTCTCAGAATGACTTG
GTTGAGTACTCACCAGTCACAGAAAAGCATCTTACGGATGGCATGACAGTAAGAGAATTA
TGCAGTGCTGCCATAACCATGAGTGATAACACTGCGGCCAACTTACTTCTGACAACGATC
GGAGGACCGAAGGAGCTAACCGCTTTTTTGCACAACATGGGGGATCATGTAACTCGCCTT
GATCGTTGGGAACCGGAGCTGAATGAAGCCATACCAAACGACGAGCGTGACACCACGATG
CCTGTAGCAATGGCAACAACGTTGCGCAAACTATTAACTGGCGAACTACTTACTCTAGCT
TCCCGGCAACAATTAATAGACTGGATGGAGGCGGATAAAGTTGCAGGACCACTTCTGCGC
TCGGCCCTTCCGGCTGGCTGGTTTATTGCTGATAAATCTGGAGCCGGTGAGCGTGGGTCT
CGCGGTATCATTGCAGCACTGGGGCCAGATGGTAAGCCCTCCCGTATCGTAGTTATCTAC
ACGACGGGGAGTCAGGCAACTATGGATGAACGAAATAGACAGATCGCTGAGATAGGTGCC
TCACTGATTAAGCATTGGTAACTGTCAGACCAAGTTTACTCATATATACTTTAGATTGAT
TTAAAACTTCATTTTTAATTTAAAAGGATCTAGGTGAAGATCCTTTTTGATAATCTCATG
ACCAAAATCCCTTAACGTGAGTTTTCGTTCCACTGAGCGTCAGACCCCGTAGAAAAGATC
AAAGGATCTTCTTGAGATCCTTTTTTTCTGCGCGTAATCTGCTGCTTGCAAACAAAAAAA
CCACCGCTACCAGCGGTGGTTTGTTTGCCGGATCAAGAGCTACCAACTCTTTTTCCGAAG
GTAACTGGCTTCAGCAGAGCGCAGATACCAAATACTGTCCTTCTAGTGTAGCCGTAGTTA
GGCCACCACTTCAAGAACTCTGTAGCACCGCCTACATACCTCGCTCTGCTAATCCTGTTA
CCAGTGGCTGCTGCCAGTGGCGATAAGTCGTGTCTTACCGGGTTGGACTCAAGACGATAG
TTACCGGATAAGGCGCAGCGGTCGGGCTGAACGGGGGGTTCGTGCACACAGCCCAGCTTG
GAGCGAACGACCTACACCGAACTGAGATACCTACAGCGTGAGCATTGAGAAAGCGCCACG
CTTCCCGAAGGGAGAAAGGCGGACAGGTATCCGGTAAGCGGCAGGGTCGGAACAGGAGAG
CGCACGAGGGAGCTTCCAGGGGGAAACGCCTGGTATCTTTATAGTCCTGTCGGGTTTCGC
CACCTCTGACTTGAGCGTCGATTTTTGTGATGCTCGTCAGGGGGGCGGAGCCTATGGAAA
AACGCCAGCAACGCGGCCTTTTTACGGTTCCTGGCCTTTTGCTGGCCTTTTGCTCACATG
TTCTTTCCTGCGTTATCCCCTGATTCTGTGGATAACCGTATTACCGCCTTTGAGTGAGCT
GATACCGCTCGCCGCAGCCGAACGACCGAGCGCAGCGAGTCAGTGAGCGAGGAAGCGGAA
GAGCGCCCAATACGCAAACCGCCTCTCCCCGCGCGTTGGCCGATTCATTAATGCAGCTGG
CACGACAGGTTTCCCGACTGGAAAGCGGGCAGTGAGCGCAACGCAATTAATGTGAGTTAG
CTCACTCATTAGGCACCCCAGGCTTTACACTTTATGCTTCCGGCTCGTATGTTGTGTGGA
ATTGTGAGCGGATAACAATTTCACACAGGAAACAGCTATGACCATGATTACGCCAAGCTT
CCGCGGGTATATGGTAAAGAACCTACTAACACAATAAAATATTTAAATAATGTATTTCCT
ATAAATAAATTTACAGATTTATTTTTTAATACAAAAGATATAGATATACCAGAAATAAAT
GATCAGTTTAAAGGTTTTAAATTCTTTATGACATCATTTATAAATCATGGATCATATCCA
CTAACAATAGAATGTGGTGTAACAAATGGTGGAACTAGTTATAAAAGAGCAATTATTTTA
TTGCATGTTCGAACTGATTTAAAAGATAGACCAGTTTCATTTTGTGATTTTCGAAAAGGA
GAATTATATAATTATTTGAATGCTTATACTGAAGGGGATGTATGCATAATAATTTCCAAA
TCAAATACAAGTTTTGGTTTTAGATGCCCAGTAAATACAAAAAAAATGCCAAAAAATTGT
TTTACGCAAGTATATGAAAAAGGGTATCTAAATGACGCCAATAAAATTAATACTAAAAAT
GTTATTAACTATTCATTTGAAAATCCAGAATATGCGCTAGCTGGTTYTAATTATACATTA
ACAAAATCGTATCAATTTGAATGTCATTGTGTAGATAAAGAAACAGAACAAATTGTAAAA
ACGGTTTTAGTCAAATATGTAAATGAAGATGAAATATATGATTATAATGATTTTCCAATG
GTGAATCACAAACCTATTATTGCACATCCAAATAAAACACATCAAGCTTGCATGCCTGCA
GGAATTCGATATCGAATTCCCATGGAACCAAAATAACATATTAATGAATTTTCTTTTATT
GTGAAAATGTTCTTATTTGTTTTAAAAAATCAAAGTAAATTTAATTCCTTTAATTGTGCT
AAATATTGTGTTTGTATAACTATAATGACATTTCCTATGCTTTTTTAATATTATATATAA
TTTTTTGTGTTTTTTCAAAAAAAACGTGTATTTGTTTTTATTTGAGTGTACAATTTGCAT
AGTGAGTTTAAAGTGTTTGAAAATTATTTAATTTATAATTTTCATAAAACGTTTAATCAT
TTATCAAAATTGCGGTTGCAATATAGACAGAAAATATTAACTTAATTTTCATTGTTTTGT
AAAACATTTTCATTATTTTTTTTAACAAAATTAAAGTAAATGAGATGGTAAATTATTTTA
ATTATAACCATTTGTTTAATTTTTTTTATTTTATTACTATTATTTTTTTTCTTTTGTGCT
GCGCAATTAAAAAGAGAATTAAGGGTGCACCAATATATTTAAGCAATCTTTTGCGACACA
CAAGAAAAAATATAACATCAACATATTTAAAATAATGGAAATTTTTTAATAATTTATTCT
ATATATATTTAATATATGTAAAAAAACTAATATTGTAATATAATTAAATAAAACTTGAAA
TTGATATAATATGTATATATATTGCATGTTTAGCTTTAAATAATATTTAAGTATTGATTT
TCTTTACACACAAAAACATTTGCCTTTTTATGTGAGGATTATTGTTTCTTTATTTATATT
GCTATGAATTTATTCTTAGTATAAATTTATTTTTTTTCGGCTTTGAAAATAATTTTATAT
ATATAAAAAGAAAAAATATGGAAAATAAAATTCATAAATCATATTTGACACATTTCAATT
TTTTTTTCTTATTTCACAGCTAGCCATATAATAATATATATGTGTCTATGGGTTCAAATA
GCTTAATTTCAATGTGCGTGATTTTTTTTTATAAAAAAACTAATTTTATGTTTGGTTATG
CATGCAACTGTTATAAAATAAAAATTGTAAAATTTGTTTTTTTGTACACATATTTGTACT
TTTTTTAATGCACATGAAAAAAGAAAGGAAAAAAAAGGGGGAAAAAAAAGGAAAAAAAAA
GGGAAAAAAGGGAAAAAACGTACATCTACGCATTGTTATTTAGCAATAAAATAGAGAACA
AATCTATAAATATATAATGTGTACAACAAATTAATTTAATTTATATTTTTACTGTAATTT
CATAATAATTTTCATTTTTACATTTTAATGTTTTTTTAATTGGAATATAATTCAAAATGA
TTTTGACACATTTTACTTATAGAAGAAGGCTATATAATTATATAAGTGTGTATATATAAG
TATTGTATGGTAATTGTTTTATAAATTAAATTTTTAAAAAACATTATTCTATTTTATAAT
TTGTAAATTAAAAATATTAATAAAATAAAACGATATAAAAGGATCCATGAGTAAAGGAGA
AGAACTTTTCACTGGAGTTGTCCCAATTCTTGTTGAATTAGATGGTGATGTTAATGGGCA
CAAATTTTCTGTCAGTGGAGAGGGTGAAGGTGATGCAACATACGGAAAACTTACCCTTAA
ATTTATTTGCACTACTGGAAAACTACCTGTTCCATGGCCAACACTTGTCACTACTTTCGG
TTATGGTGTTCAATGCTTTGCGAGATACCCAGATCATATGAAACAGCATGACTTTTTCAA
GAGTGCCATGCCCGAAGGTTATGTACAGGAAAGAACTATATTTTTCAAAGATGACGGGAA
CTACAAGACACGTGCTGAAGTCAAGTTTGAAGGTGATACCCTTGTTAATAGAATCGAGTT
AAAAGGTATTGATTTTAAAGAAGATGGAAACATTCTTGGACACAAATTGGAATACAACTA
TAACTCACACAATGTATACATCATGGCAGACAAACAAAAGAATGGAATCAAAGTTAACTT
CAAAATTAGACACAACATTGAAGATGGAAGCGTTCAACTAGCAGACCATTATCAACAAAA
TACTCCAATTGGCGATGGCCCTGTCCTTTTACCAGACAACCATTACCTGTCCACACAATC
TGCCCTTTCGAAAGATCCCAACGAAAAGAGAGACCACATGGTCCTTCTTGAGTTTGTAAC
AGCTGCTGGGATTACACATGGCATGGATGAACTATACAAAGGGATCCTGGCTAGCCAGTC
GACCTGCAGGCATGCAAGCTTGCGGCCGATCCAAATGGAAGACGCCAAAAACATAAAGAA
AGGCCCGGCGCCATTCTATCCGCTGGAAGATGGAACCGCTGGAGAGCAACTGCATAAGGC
TATGAAGAGATACGCCCTGGTTCCTGGAACAATTGCTTTTACAGATGCACATATCGAGGT
GAACATCACGTACGCGGAATACTTCGAAATGTCCGTTCGGTTGGCAGAAGCTATGAAACG
ATATGGGCTGAATACAAATCACAGAATCGTCGTATGCAGTGAAAACTCTCTTCAATTCTT
TATGCCGGTGTTGGGCGCGTTATTTATCGGAGTTGCAGTTGCGCCCGCGAACGACATTTA
TAATGAACGTGAATTGCTCAACAGTATGAACATTTCGCAGCCTACCGTAGTGTTTGTTTC
CAAAAAGGGGTTGCAAAAAATTTTGAACGTGCAAAAAAAATTACCAATAATCCAGAAAAT
TATTATCATGGATTCTAAAACGGATTACCAGGGATTTCAGTCGATGTACACGTTCGTCAC
ATCTCATCTACCTCCCGGTTTTAATGAATACGATTTTGTACCAGAGTCCTTTGATCGTGA
CAAAACAATTGCACTGATAATGAATTCCTCTGGATCTACTGGGTTACCTAAGGGTGTGGC
CCTTCCGCATAGAACTGCCTGCGTCAGATTCTCGCATGCCAGAGATCCTATTTTTGGCAA
TCAAATCATTCCGGATACTGCGATTTTAAGTGTTGTTCCATTCCATCACGGTTTTGGAAT
GTTTACTACACTCGGATATTTGATATGTGGATTTCGAGTCGTCTTAATGTATAGATTTGA
AGAAGAGCTGTTTTTACGATCCCTTCAGGATTACAAAATTCAAAGTGCGTTGCTAGTACC
AACCCTATTTTCATTCTTCGCCAAAAGCACTCTGATTGACAAATACGATTTATCTAATTT
ACACGAAATTGCTTCTGGGGGCGCACCTCTTTCGAAAGAAGTCGGGGAAGCGGTTGCAAA
ACGCTTCCATCTTCCAGGGATACGACAAGGATATGGGCTCACTGAGACTACATCAGCTAT
TCTGATTACACCCGAGGGGGATGATAAACCGGGCGCGGTCGGTAAAGTTGTTCCATTTTT
TGAAGCGAAGGTTGTGGATCTGGATACCGGGAAAACGCTGGGCGTTAATCAGAGAGGCGA
ATTATGTGTCAGAGGACCTATGATTATGTCCGGTTATGTAAACAATCCGGAAGCGACCAA
CGCCTTGATTGACAAGGATGGATGGCTACATTCTGGAGACATAGCTTACTGGGACGAAGA
CGAACACTTCTTCATAGTTGACCGCTTGAAGTCTTTAATTAAATACAAAGGATATCAGGT
GGCCCCCGCTGAATTGGAATCGATATTGTTACAACACCCCAACATCTTCGACGCGGGCGT
GGCAGGTCTTCCCGACGATGACGCCGGTGAACTTCCCGCCGCCGTTGTTGTTTTGGAGCA
CGGAAAGACGATGACGGAAAAAGAGATCGTGGATTACGTCGCCAGTCAAGTAACAACCGC
GAAAAAGTTGCGCGGAGGAGTTGTGTTTGTGGACGAAGTACCGAAAGGTCTTACCGGAAA
ACTCGACGCAAGAAAAATCAGAGAGATCCTCATAAAGGCCAAGAAGGGCGGAAAGATCGC
CGTGTAATTCTAGAAGATCCCGTTTTTCTTACTTATATATTTATACCAATTGATTGTATT
TATAACTGTAAAAATGTGTATGTTGTGTGCATATTTTTTTTTGTGCATGCACATGCATGT
AAATAGCTAAAATTATGAACATTTTATTTTTTGTTCAGAAAAAAAAAACTTTACACACAT
AAAATGGCTAGTATGAATAGCCATATTTTATATAAATTAAATCCTATGAATTTATGACCA
TATTAAAAATTTAGATATTTATGGAACATAATATGTTTGAAACAATAAGACAAAATTATT
ATTATTATTATTATTTTTACTGTTATAATTATGTTGTCTCTTCAATGATTCATAAATAGT
TGGACTTGATTTTTAAAATGTTTATAATATGATTAGCATAGTTAAATAAAAAAAGTTGAA
AAATTAAAAAAAAACATATAAACACAAATGATGTTTTTTCCTTCAATTTCGGGTACCGAG
CTCGAATTCTCTTGAGCCCGTTAATGAAATAGATACAATTCATTCATGTTATATACATCT
AGAACATAATCTGAATATGGTTCAAGTTAAATGTCCAAAAATTATAAAAAGTGATGATAT
TTTTGATGGTAATACCATAATAGACACCAAGGTAACATCACGAAGTAGTCAACAAAATAA
TTTTTATTTAGAAAATACAGATGTTGAACCAGAAGAAATAGAGAAATATAAAAATATAGA
ATACATACCAGAAAACGATGAAGTAATGCATCTAGACAAAAAAGAAAAGCTAGATGATAT
ATTACCAGGTGTTATCATATTAGATAAAAATAAAATGTTCAAAGAAAAAGGACATTTCAC
TTTTGTTACTCCATTAATTGTAGAAAAGGTATTAATATTAAAAATATATTGTGATAATAC
TAAAACAATAATTAATAATATGAAAGGGAAAAAAGGTATTACAGTAATAAGGATTTCTCA
AAATACAACAAAAAATAAATTTTATGGATGTGACTTTTCAGGTAATTCTAAAAAAACATT
TTACTATTCCAATGTTTATGATTTAGAAAAAAAAAATGAGTTTTGTGAAATAGAATTAAA
AGAAAATATAGTAGTTAGCTTAAATTGTCCAACTGGTAAAATTAATCCAAAAAATTGTTT
TAGAAATGTATATATAAAAAGTAATATGAATGAACAAACAACCGAAAATATAGAAAATAT
ATTTAACGAAATAAAAGTTATAGATGCAGATTATTTTATAAATAATTCATCAACCTTTTT
GATGATTTCCAAAATTACAAAAAAAGAGTTTGATTTTTATTGTACATGTGAAGATTATAA
AACCAAAAATATAGGAACAATATATATTAAAAATTATGAATATCTAGATTCAAAACCTAA
ATATAAAAATAAACAAATTTCCTATATAGATGTAGTTCCATACCCGCGGGGAAAGGGCG
Restriction sites to linearize plasmid KspI (SacII)
Selectable marker used to select the mutant parasitegfp (FACS)
Promoter of the selectable markerama-1
Selection (positive) procedureFACS (flowsorting)
Selection (negative) procedureNo
Additional remarks genetic modificationThe GFP-Luciferase gene (1 copy) has been inserted into the 230p locus (PB000214.00.0) by double cross-over integration
Additional remarks selection procedureThis reporter mutant expressing GFP-Luciferase does not contain a drug-selectable marker. This mutant has been selected by FACS sorting after transfection based on GFP fluorescence
Other details transgene
Promoter
Gene Model of Parasite PBANKA_0915000
Gene Model P. falciparum ortholog PF3D7_1133400
Gene productapical membrane antigen 1
Gene product: Alternative nameAMA-1
Primer information details of the primers used for amplification of the promoter sequence  Click to view information
Primer information details of the primers used for amplification of the promoter sequence  Click to hide information
Sequence Primer 1
Additional information primer 1
Sequence Primer 2
Additional information primer 2
3'-UTR
Gene Model of Parasite PBANKA_0719300
Gene productbifunctional dihydrofolate reductase-thymidylate synthase, putative
Gene product: Alternative namedhfr/ts
Primer information details of the primers used for amplification the 3'-UTR sequences  Click to view information
Primer information details of the primers used for amplification the 3'-UTR sequences  Click to hide information
Sequence Primer 1
Additional information primer 1
Sequence Primer 2
Additional information primer 2
Insertion/Replacement locus
Replacement / InsertionReplacement locus
Gene Model of Parasite PBANKA_0306000
Gene product6-cysteine protein
Gene product: Alternative name230p
Primer information details of the primers used for amplification of the target sequences  Click to view information
Primer information details of the 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