Summary

RMgm-130
Malaria parasiteP. berghei
Genotype
Transgene
Transgene not Plasmodium: GFP (gfp-mut3)
Promoter: Gene model: PBANKA_0416100; Gene model (P.falciparum): PF3D7_0905300; Gene product: dynein heavy chain, putative
3'UTR: Gene model: PBANKA_0719300; Gene product: bifunctional dihydrofolate reductase-thymidylate synthase, putative (dhfr/ts)
Insertion locus: Gene model: Not available; Gene product: Not available (small subunit ribosomal rna gene (c- or d-type unit))
Phenotype Gametocyte/Gamete;
Last modified: 18 February 2009, 13:52
  *RMgm-130
Successful modificationThe parasite was generated by the genetic modification
The mutant contains the following genetic modification(s) Introduction of a transgene
Reference (PubMed-PMID number) Reference 1 (PMID number) : 15935755
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/ResearcherS.M. Khan, G.R. Mair, C.J. Janse, A.P. Waters
Name Group/DepartmentLeiden Malaria Research Group
Name InstituteLeiden University Medical Center (LUMC)
CityLeiden
CountryThe Netherlands
Name of the mutant parasite
RMgm numberRMgm-130
Principal name520
Alternative name
Standardized name
Is the mutant parasite cloned after genetic modificationNo
Phenotype
Asexual blood stageNot different from wild type
Gametocyte/GameteGFP expression in male gametocytes. No expression could be detected in asexual blood stages and female gametocytes.
Fertilization and ookineteNot tested
OocystNot tested
SporozoiteNot tested
Liver stageNot tested
Additional remarks phenotype

Mutant/mutation
The mutant expresses GFP under the control of the promoter of the gene encoding PB000791.03.0 (PFI0260c; dynein heavy chain, putative).

Protein (function)
PB000791.03.0 (PFI0260c; dynein heavy chain, putative).

Phenotype
Phenotype analyses indicate absence of promoter activity in asexual blood stages and female gametocytes and active transcription in male gametocytes. The absence in female gametocytes and presence in males is in agreement with proteome analysis of males and females (Khan, S.M. et al., 2005, Cell, 121, 675-87). See further 'Additional information.

Additional information
The construct GFPcon (RMgm-5) containing GFP under the control of the eef1a promoter formed the basis for the construct driving expression of GFP under control of the PB000791.03.0 promoter. This constructs contains a c/d-ssurrna target fragment for integration into the c/d-rrna genomic locus. GFP is flanked by the 3'-UTR sequence of the dhfr gene of P. berghei. The eef1a promoter region is replaced by a 1527 bp fragment (EcoRV/BamHI) of the promoter region of PB000791.03.0.
Because it has been shown that 3'-UTR regions can influence expression of genes in Plasmodium, care has to be taken in interpretation of expression of transgenes when 3'-UTR sequences of heterologous genes are used.

Other mutants


  Transgene: Mutant parasite expressing a transgene
Type and details of transgene
Is the transgene Plasmodium derived Transgene: not Plasmodium
Transgene nameGFP (gfp-mut3)
Details of the genetic modification
Inducable system usedNo
Additional remarks inducable system
Type of plasmid/constructPlasmid single 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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ATCATATTTAAACAGATTAAGTACCGAAGATAATAATGATGATGATGAAACAAAAGGTTC
TTATACTAATGAGGAAGATAACGCCCCTGCATCTGAACCTTATACCGATGAAGATAAAAA
TTTATTATAATTTAAAAGCTTCATATTTTATTATACTATGTATGTATTATATGTACAAAG
ACATGTAGGCAAGTCATTGGAAAAGTATTTTGGTGTTTAATGTATTTATATGTGCATGCT
TGCACCTACTTTTCTATTAATTTTCCGAGCTTTATAATTTCTTGTCAATAAAATATAAAA
ACATATATACACATGATAAATTTTTCCTTTTCATAAATCCTTCAAAACTGTCAAATAAAT
AATTATAATGTATATATATGTTTTATTTATACAAAATTATGTGCATATATATATTTACAT
TTAAAACCGAATGAATTTTTATATAAACCCTCATAAAATAAGCTTTTAAATTTAATATAA
TAAAGGGGCTTGTGAGTTAAATAAAATGGTAACACATTTTTAAATAGTACCTTTTAAATT
ATTTCTGTATCTAGTATAAATAATATTAAAGGTAATAAGAAATAGGAAAAAAATTTTATA
TATATAAAATGGGGACGTGAAATAAAATTAACATGTTCATATAAAAATTGTGGTTCGAAA
ATTTGGGTATAAACGCATTTAAAATAGGTACATAATATATTTTATTCGTTTTGGGTATTT
ATTTTTTTATGGTTAGTATATAATTATAAAATTGTTTTTTTTCATTCAAAGTTTGTAAGT
AAAATGTTGAGATGTTTTAGAAACAATAATAAGCTTCGAAATGCTAAACTTTGAGATAAG
ATGTGGGTTTGTCTTAAGACACGATTAGAAGCAGTTCGAACCGTTTCTAAATAATTAAGA
AATGAATGATAATATGCGTATTTATTTGCATTTTTGCGTATTCACAATCTTATTTATGCA
CATTTAAATTTTATTGTTTTGAGACTTCAAAGAGCTTATTCACCACTACTAGTTAATAAT
ATATATTTTCTCCTAAGTTTATTATAAATGGGAATTTTCGACTTTTACATGTATAAAAAA
AAAAAATTGGAGAAAAAAAGGGGATATAATATGCAATAAAATAAGTAGATATGTACACAT
ACTATATATTTATATATGTGTGTACGTATATGGGAAAGTTATATTAAAGAAAATATATTA
TAAAATGTTTATGCAAGATAATATAAATATAAATATAAATAAATGGATATATATTTTCAT
ATGACAGTATATATATATATATATATATATATTCATTTTTATATGCGAAAAAGGGGTTTG
TGCATAAGGTTTATAGTATATATAGATATTTGCAAATATATGGACAAACATATATATGCA
GTGTTAAGGGCACAAGTTATTTGGATATGTAATTTGGTGGTATATAGTAGTATTATATGT
ATTTCATACATATAGATGCATGTACATATGAATAGGAAGTTTGATTTAATTCAGAATAAA
TATAAGAATTAATTATCCAAATGATAAAAAGGATCCATGAGTAAAGGAGAAGAACTTTTC
ACTGGAGTTGTCCCAATTCTTGTTGAATTAGATGGTGATGTTAATGGGCACAAATTTTCT
GTCAGTGGAGAGGGTGAAGGTGATGCAACATACGGAAAACTTACCCTTAAATTTATTTGC
ACTACTGGAAAACTACCTGTTCCATGGCCAACACTTGTCACTACTTTCGGTTATGGTGTT
CAATGCTTTGCGAGATACCCAGATCATATGAAACAGCATGACTTTTTCAAGAGTGCCATG
CCCGAAGGTTATGTACAGGAAAGAACTATATTTTTCAAAGATGACGGGAACTACAAGACA
CGTGCTGAAGTCAAGTTTGAAGGTGATACCCTTGTTAATAGAATCGAGTTAAAAGGTATT
GATTTTAAAGAAGATGGAAACATTCTTGGACACAAATTGGAATACAACTATAACTCACAC
AATGTATACATCATGGCAGACAAACAAAAGAATGGAATCAAAGTTAACTTCAAAATTAGA
CACAACATTGAAGATGGAAGCGTTCAACTAGCAGACCATTATCAACAAAATACTCCAATT
GGCGATGGCCCTGTCCTTTTACCAGACAACCATTACCTGTCCACACAATCTGCCCTTTCG
AAAGATCCCAACGAAAAGAGAGACCACATGGTCCTTCTTGAGTTTGTAACAGCTGCTGGG
ATTACACATGGCATGGATGAACTATACAAATAAATTCTAGAAGATCCCGTTTTTCTTACT
TATATATTTATACCAATTGATTGTATTTATAACTGTAAAAATGTGTATGTTGTGTGCATA
TTTTTTTTTGTGCATGCACATGCATGTAAATAGCTAAAATTATGAACATTTTATTTTTTG
TTCAGAAAAAAAAAACTTTACACACATAAAATGGCTAGTATGAATAGCCATATTTTATAT
AAATTAAATCCTATGAATTTATGACCATATTAAAAATTTAGATATTTATGGAACATAATA
TGTTTGAAACAATAAGACAAAATTATTATTATTATTATTATTTTTACTGTTATAATTATG
TTGTCTCTTCAATGATTCATAAATAGTTGGACTTGATTTTTAAAATGTTTATAATATGAT
TAGCATAGTTAAATAAAAAAAGTTGAAAAATTAAAAAAAAACATATAAACACAAATGATG
TTTTTTCCTTCAATTTCGGGTACCGAGCTCGAATTCAACCTGGTTGATCTTGCCAGTAGT
CATATGCTTGTCTCAAAGATTAAGCCATGCAAGTGAAAGTATATGCACATTTATTGCAGA
AACTGCGAACGGCTCATTAAAACAGTTATAATCTACTTGACATTTTATTATAAGGATAAC
TACGGAAAAGCTGTAGCTAATACTTGTTAAGTACTTTTACTCCCCGGAGTAATTGTATGT
ATTTGTTAAGACCCCTAAGAAAAAATGATATTAAAGGAATTATAACAAAGAAGCAACACA
TAATATAATTATTCAGTGTGTATCAATCGAGTTTCTGACCTATCAGCTTTTGATGTTAGG
GTATTGACCTAACATGGCTTTGACGGGTAACGGGGAATTAGAGTTCGATTCCGGAGAGGG
AGCCTGAGAAATAGCTACCACATCTAAGGAAGGCAGCAGGCGCGTAAATTACCCAATTCT
AAATAAGAGAGGTAGTGACAAGAAATAACAATATAAGGCCAAATTTTGGTTTTATAATTG
GAATGATGGGAATTTAAAACCTTCCCAAAAATCAATTGGAGGGCAAGTCTGGTGCCAGCA
GCCGCGGTAATTCCAGCTCCAATAGCGTATATTAAAATTGTTGCAGTTAAAACGCTCGTA
GTTGAACTTCAAGGGTATAATTATTTTAAGCAACTCACTTGGAAAGAATCATGACTTCTG
TCACTGCTTTTATCCTTGTTGCAGTTCTTTTAATACAGGGCCCTTTGAGAGCCCATTAAT
TTATGACTGGGTTTCTCGTTACTTTGAGTAAATTAGAGTGTTTAAAGCAAACAGATAAAG
CGTATTTTACTGTGTTTGAATACTATAGCATGGAATAACAACATTGAATAGGTCAAAAGT
TTTTGAAAAATTTTTCTTATTTTGGCTTAGATACAGTTAATAGGAGTAGCTTGGGGGCAT
TTGTATTCAGATGTCAGAGGTGAAATTCTTAGATTTTCTGGAGACAAACAACTGCGAAAG
CATTTGCCTAAAATACTTCCATTAATCAAGAACGAAAGTTAAGGGAGTGAAGACGATCAG
ATACCGTCGTAATCTTAACCATAAACTATGCCGACTAAGTGTTGGATGAAAATTTATAAA
TAAAACTATCTTCTTTAAAGGAGTAGTTTTTTAGATGCTTCCTTCAGTACCTTATGAGAA
ATCAAAGTCTTTGGGTTCTGGGGCGAGTATTCGCGCAAGCGAGAAAGTTAAAAGAATTGA
CGGAAGGGCACCACCAGGCGTGGAGCTTGCGGCTTAATTTGACTCAACACGGGGAAACTC
ACTAGTTTAAGACAAGAGTAGGATTGACAGATTAATAGCTCTTTCTTGATTTCTTGGATG
GTGATGCATGGCCGTTTTTAGTTCGTGAATATGATTTGTCTGGTTAATTCCGATAACGAA
CGAGATCTTAACCTGCTAATTAGCGGTGGATATGTGATATTCTTCGAAGGTGGACTAACT
ATAGCGTTTTCGAAGGTATGTTGCATAATCAAATTGGTTTACCCTTTGTTTTTTTGTAGC
ATATTCTTTTATTTCGTTGGGTTTTTTCCCTAGTAAGGATGTATCTGCTTTATTTAATGC
TTCTTAGAGGAACGATGTGTGTCTAACACAAGGAAGTTTAAGGCAACAACAGGTCTGTGA
TGTCCTTAGATATACTAGGCTGCACGCGTGCTACACTGATATGTAAAACGAGTATTTAAA
ATTATATCTGTATGGTAGATAATTTAATTTCTACGTATTATCAGCATATACTTTTCCTAC
ACTGAAATAGTGAAGGTAATCTTTATCAATACATATCGTGATGGGGATAGATTATTGCAA
TTATTAATCTTGAACGAGGAATGCCTAGTAAGCATGATTCATCAGATTGTGCTGACTACG
TCCCTGCCCTTTGTACACACCGCCCGTCGCTCCTACCGATTGAAAGATATGATGAATTGT
TTGGACAAGAAAATAGAAATTTTATTTTTATTTTTTTTGGAAGGACCGTAAATCCTATCT
TTTAAAGGAAGGAGAAGTCGTAACAAGGTTTCCGTAGGTGAATTCACTGGCCGTCGTTTT
ACAACGTCGTGACTGGGAAAACCCTGGCGTTACCCAACTTAATCGCCTTGCAGCACATCC
CCCTTTCGCCAGCTGGCGTAATAGCGAAGAGGCCCGCACCGATCGCCCTTCCCAACAGTT
GCGCAGCCTGAATGGCGAATGGCGCCTGATGCGGTATTTTCTCCTTACGCATCTGTGCGG
TATTTCACACCGCATATGGTGCACTCTCAGTACAATCTGCTCTGATGCCGCATAGTTAAG
CCAGCCCCGACACCCGCCAACACCCGCTGACGCGCCCTGACGGGCTTGTCTGCTCCCGGC
ATCCGCTTACAGACAAGCTGTGACCGTCTCCGGGAGCTGCATGTGTCAGAGGTTTTCACC
GTCATCACCGAAACGCGCGAGACGAAAGGGCCTCGTGATACGCCTATTTTTATAGGTTAA
TGTCATGATAATAATGGTTTCTTAGACGTCAGGTGGCACTTTTCGGGGAAATGTGCGCGG
AACCCCTATTTGTTTATTTTTCTAAATACATTCAAATATGTATCCGCTCATGAGACAATA
ACCCTGATAAATGCTTCAATAATATTGAAAAAGGAAGAGTATGAGTATTCAACATTTCCG
TGTCGCCCTTATTCCCTTTTTTGCGGCATTTTGCCTTCCTGTTTTTGCTCACCCAGAAAC
GCTGGTGAAAGTAAAAGATGCTGAAGATCAGTTGGGTGCACGAGTGGGTTACATCGAACT
GGATCTCAACAGCGGTAAGATCCTTGAGAGTTTTCGCCCCGAAGAACGTTTTCCAATGAT
GAGCACTTTTAAAGTTCTGCTATGTGGCGCGGTATTATCCCGTATTGACGCCGGGCAAGA
GCAACTCGGTCGCCGCATACACTATTCTCAGAATGACTTGGTTGAGTACTCACCAGTCAC
AGAAAAGCATCTTACGGATGGCATGACAGTAAGAGAATTATGCAGTGCTGCCATAACCAT
GAGTGATAACACTGCGGCCAACTTACTTCTGACAACGATCGGAGGACCGAAGGAGCTAAC
CGCTTTTTTGCACAACATGGGGGATCATGTAACTCGCCTTGATCGTTGGGAACCGGAGCT
GAATGAAGCCATACCAAACGACGAGCGTGACACCACGATGCCTGTAGCAATGGCAACAAC
GTTGCGCAAACTATTAACTGGCGAACTACTTACTCTAGCTTCCCGGCAACAATTAATAGA
CTGGATGGAGGCGGATAAAGTTGCAGGACCACTTCTGCGCTCGGCCCTTCCGGCTGGCTG
GTTTATTGCTGATAAATCTGGAGCCGGTGAGCGTGGGTCTCGCGGTATCATTGCAGCACT
GGGGCCAGATGGTAAGCCCTCCCGTATCGTAGTTATCTACACGACGGGGAGTCAGGCAAC
TATGGATGAACGAAATAGACAGATCGCTGAGATAGGTGCCTCACTGATTAAGCATTGGTA
ACTGTCAGACCAAGTTTACTCATATATACTTTAGATTGATTTAAAACTTCATTTTTAATT
TAAAAGGATCTAGGTGAAGATCCTTTTTGATAATCTCATGACCAAAATCCCTTAACGTGA
GTTTTCGTTCCACTGAGCGTCAGACCCCGTAGAAAAGATCAAAGGATCTTCTTGAGATCC
TTTTTTTCTGCGCGTAATCTGCTGCTTGCAAACAAAAAAACCACCGCTACCAGCGGTGGT
TTGTTTGCCGGATCAAGAGCTACCAACTCTTTTTCCGAAGGTAACTGGCTTCAGCAGAGC
GCAGATACCAAATACTGTCCTTCTAGTGTAGCCGTAGTTAGGCCACCACTTCAAGAACTC
TGTAGCACCGCCTACATACCTCGCTCTGCTAATCCTGTTACCAGTGGCTGCTGCCAGTGG
CGATAAGTCGTGTCTTACCGGGTTGGACTCAAGACGATAGTTACCGGATAAGGCGCAGCG
GTCGGGCTGAACGGGGGGTTCGTGCACACAGCCCAGCTTGGAGCGAACGACCTACACCGA
ACTGAGATACCTACAGCGTGAGCATTGAGAAAGCGCCACGCTTCCCGAAGGGAGAAAGGC
GGACAGGTATCCGGTAAGCGGCAGGGTCGGAACAGGAGAGCGCACGAGGGAGCTTCCAGG
GGGAAACGCCTGGTATCTTTATAGTCCTGTCGGGTTTCGCCACCTCTGACTTGAGCGTCG
ATTTTTGTGATGCTCGTCAGGGGGGCGGAGCCTATGGAAAAACGCCAGCAACGCGGCCTT
TTTACGGTTCCTGGCCTTTTGCTGGCCTTTTGCTCACATGTTCTTTCCTGCGTTATCCCC
TGATTCTGTGGATAACCGTATTACCGCCTTTGAGTGAGCTGATACCGCTCGCCGCAGCCG
AACGACCGAGCGCAGCGAGTCAGTGAGCGAGGAAGCGGAAGAGCGCCCAATACGCAAACC
GCCTCTCCCCGCGCGTTGGCCGATTCATTAATGCAGCTGGCACGACAGGTTTCCCGACTG
GAAAGCGGGCAGTGAGCGCAACGCAATTAATGTGAGTTAGCTCACTCATTAGGCACCCCA
GGCTTTACACTTTATGCTTCCGGCTCGTATGTTGTGTGGAATTGTGAGCGGATAACAATT
TCACACAGGAAACAGCTATGACCATGATTACGCCAAGCTTGCATGCCTGCAGGTCAACAA
TAAATAATAAATAAATATTGTGGAAATAAAATAACATATAATTATTTTTAATACATTGAT
TTCCCTTTTATTTTTTTAAATTTCATTGATATAAAAATATATAATAATAACATATATGAT
TTCAAATTAATCTTTTCAAAAATGGTGTTTATTTTTGTATGTTTGTGTATGAATTAATCA
CATAACACATCTATTAAATTGAGTTGGTAATATAGACACAAATAAATATATATATTTTTA
TAGCTTAAAAGTGTGTTATGAATATTTTAAGCATATTTTCTTTTTCTTTGGATTGTGTAA
AATGAACTCATATAATGCGTTTTTTTGTTTTTGTTATTTTGTCATTTTGTTATTTTGCTA
TTTTATGGATTAATTTTTGTTTATAAAATGGGAAATAATTTTAACATATTTAAATAAATG
GAGAAAAAATATAAAATAATTATAAAAAAAAGTTAATACACATTTTTTCCTGTTATAGAC
CTTATATTTATTTATCCATATATATATATATATATATATATATATATATATACATACCAA
GTGAATTAAGAGGAAAGCTAATTTATTATTCAGAATAATATATGAACTATATATAATTTT
TATTATTTTGGTGTATATTAATCTGTCTATATGCATACATGCAATAATTTATCGACTTAT
ATATCAAATAACATAAAATAGAAGTGTTTTAAATTATGGATATATGCTCAATATTCATTT
TTTTTAATAAGTTAGCTATATTTAAATTATACATTTTATATATGGTCTCTTTTTTTTTTA
AATATTATTTAAGTGATCATGAAAATATAAATAATTTTTTTTTATTTAATATCCTTTTGC
TTGCATGTGGTAAATGGAAATTTGGATGTGTTTTGAARGTTCGGATATAGTTGTATGGAC
ATATAATATATTTTGTGAAAAATTGGTTTTATGTTTATACTTATGCCAATACTTTTTGAG
TAAAACAAAGCAAGTGCTTATAAATAATTAAAGCCAATTTTATAATATATATTTTTTTAT
TTAATTTGAATTTAGTAGTATAATTTTTTATGGTAAGTGCTCAAAGAGAGTTGCTTATAA
AGTATGGTTTGTTTCTTTTTCGCCATTTTGAATTACACATTAAAAATATATAGATACATA
TATTATAATATGAAATCATTAATAATTTAGGGAAATTCTACAAATTTAAAAACGAATAAA
ATAATTGTTTTTCATCATGCCATAACACAATATTGATATATACATGTACAAACATTTTTT
TTATTTGGAAAATATAAATTATATAAAAAAAAATGTATAGTATACAAAATGAGCATATTC
ACACGGGGTGGACGTTCATTTTTTCATTTTTCCCCTGTTTTTTATGAGTATATGATAAAA
TTTTATGAACATTTACACAAAATGAAAATGGATATATAGGAAAAATGGAGCGGTATTTCA
TTTATCTTTGATTGTCATTTGGATATTATATTACCYTGGGTAGGCAATTAAAAATGTTAA
ATAACAATTTAAGGAAATTATATTTTATATATTAAAATTAACACTGTATTATATGATTCG
CTTATAAAAGCCACTCTTTCCCCATGCAAAGCTGTTTAATATCAATTTTAACAAATTACA
CACATGTTAATATATTTATATATATAATTTATATATTTATAATTTATATATTTATATTTT
TATTATTTATATATTTATTATTTATTGTGTGTGTCAATTCGGGTAGGATATACCTCTTTT
TTATTGTTTAAAGCGATTTGTATTCTAAAATATAAAGRATTTGAAAAAGAGAAAGATAGA
ATATGATCCCATCATATATAGCCCTATAATTTTTATTTAGCAGCGAATTAATTTTTCTAT
TAAGTTTATGTGTAATTAAAATAACGGAATATATATAATACAATAAAAAAGTGCATAAAT
TAAAATTTTTTCAATTAAATTTTTTTTTTTAAGGGGTTATATAATATTAAATATATAAAA
TACGATTATATATTTTTGCTACAATTTTTTATATTAAGATATAAATAGTAAATAAATGGT
ATTATATGGCATGTAATATATAAATTTTTTCCAATTTTTATTTTATATACACTTTTCCTT
TTTTTGTCATAAAACTTAAACAATTTACACATTCATTTTAAAAATTGACTATTTGTTTCA
ACATTTTTTGAGTTTCCGTTTTATAATAGTATTTTCATTTGTATATTGCTTATATATATA
AATACACACCTAAATGTTACAAAGGATCAATGCATAAACCGGTGTGTCTGGTCGTCGCGA
TGACCCCCAAGAGGGGCATCGGCATCAACAACGGCCTCCCGTGGCCCCACTTGACCACAG
ATTTCAAACACTTTTCTCGTGTGACAAAAACGACGCCCGAAGAAGCCAGTCGCCTGAACG
GGTGGCTTCCCAGGAAATTTGCAAAGACGGGCGACTCTGGACTTCCCTCTCCATCAGTCG
GCAAGAGATTCAACGCCGTTGTCATGGGACGGAAAACCTGGGAAAGCATGCCTCGAAAGT
TTAGACCCCTCGTGGACAGATTGAACATCGTCGTTTCCTCTTCCCTCAAAGAAGAAGACA
TTGCGGCGGAGAAGCCTCAAGCTGAAGGCCAGCAGCGCGTCCGAGTCTGTGCTTCACTCC
CAGCAGCTCTCAGCCTTCTGGAGGAAGAGTACAAGGATTCTGTCGACCAGATTTTTGTCG
TGGGAGGAGCGGGACTGTACGAGGCAGCGCTGTCTCTGGGCGTTGCCTCTCACCTGTACA
TCACGCGTGTAGCCCGCGAGTTTCCGTGCGACGTTTTCTTCCCTGCGTTCCCCGGAGATG
ACATTCTTTCAAACAAATCAACTGCTGCGCAGGCTGCAGCTCCTGCCGAGTCTGTGTTCG
TTCCCTTTTGTCCGGAGCTCGGAAGAGAGAAGGACAATGAAGCGACGTATCGACCCATCT
TCATTTCCAAGACCTTCTCAGACAACGGGGTTCCCTACGACTTTGTGGTTCTCGAGAAGA
GAAGGAAGACTGACGACGCAGCCACTGCGGAACCGAGCAACGCAATGAGCTCCTTGACGT
CCACGAGGGAGACAACTCCCGTGCACGGGTTGCAGGCTCCTTCTTCGGCCGCAGCCATTG
CCCCGGTGTTGGCGTGGATGGACGAAGAAGACCGGAAAAAACGCGAGCAAAAGGAACTGA
TTCGGGCCGTTCCGCATGTTCACTTTAGAGGCCATGAAGAGTTCCAGTACCTTGATCTCA
TTGCCGACATTATTAACAATGGAAGGACAATGGATGACCGAACGGGCGTTGGTGTCATCT
CCAAATTCGGCTGCACTATGCGCTACTCGCTGGATCAGGCCTTTCCACTTCTCACCACAA
AGCGTGTGTTCTGGAAAGGGGTCCTCGAAGAGTTGCTGTGGTTCATTCGCGGCGACACGA
ACGCAAACCATCTTTCTGAGAAGGGCGTGAAGATCTGGGACAAGAATGTGACACGCGAGT
TCCTCGATTCGCGCAATCTCCCCCACCGAGAGGTCGGAGACATCGGCCCGGGCTACGGCT
TCCAGTGGAGACACTTCGGCGCGGCATACAAAGACATGCACACAGACTACACAGGGCAGG
GCGTCGACCAGCTGAAGAATGTGATCCAGATGCTGAGAACGAATCCAACAGATCGTCGCA
TGCTCATGACTGCCTGGAATCCTGCAGCGCTGGACGAAATGGCGCTGCCGCCTTGTCACT
TGTTGTGCCAGTTCTACGTGAACGACCAGAAGGAGCTGTCGTGCATCATGTATCAGCGGT
CGTGCGATGTCGGCCTCGGCGTCCCCTTCAACATCGCTTCCTATTCGCTTTTGACGCTCA
TGGTTGCACACGTCTGCAACCTAAAACCTAAGGAGTTCATTCACTTCATGGGGAACACGC
ATGTCTACACGAACCATGTCGAGGCTTTAAAAGAGCAGCTGCGGAGAGAACCGAGACCGT
TCCCCATTGTGAACATCCTCAACAAGGAACGCATCAAGGAAATCGACGATTTCACCGCCG
AGGATTTTGAGGTCGTGGGCTACGTCCCGCACGGACGAATCCAGATGGAGATGGCTGTCT
AGCGGAAATACAGAAGCTAGCTTTGATCCCGTTTTTCTTACTTATATATTTATACCAATT
GATTGTATTTATAACTGTAAAAATGTGTATGTTGTGTGCATATTTTTTTTTGTGCATGCA
CATGCATGTAAATAGCTAAAATTATGAACATTTTATTTTTTGTTCAGAAAAAAAAAACTT
TACACACATAAAATGGCTAGTATGAATAGCCATATTTTATATAAATTAAATCCTATGAAT
TTATGACCATATTAAAAATTTAGATATTTATGGAACATAATATGTTTGAAACAATAAGAC
AAAATTATTATTATTATTATTATTTTTACTGTTATAATTATGTTGTCTCTTCAATGATTC
ATAAATAGTTGGACTTGATTTTTAAAATGTTTATAATATGATTAGCATAGTTAAATAAAA
AAAGTTGAAAAATTAAAAAAAAACATATAAACACAAATGATGTTTTTTCCTTCAATTTCG
AT
Restriction sites to linearize plasmid
Selectable marker used to select the mutant parasitetgdhfr
Promoter of the selectable markerpbdhfr
Selection (positive) procedurepyrimethamine
Selection (negative) procedureNo
Additional remarks genetic modificationThe construct GFPcon (RMgm-5) containing GFP under the control of the eef1a promoter formed the basis for the construct driving expression of GFP under control of the PB000791.03.0 promoter.
This constructs contains a c/d-ssurrna target fragment for integration into the c/d-rrna genomic locus. GFP is flanked by the 3'-UTR sequence of the dhfr gene of P. berghei. The eef1a promoter region is replaced by a 1527 bp fragment (EcoRV/BamHI) of the promoter region of PB000791.03.0.

Because it has been shown that 3'-UTR regions can influence expression of genes in Plasmodium, care has to be taken in interpretation of expression of transgenes when 3'-UTR sequences of heterologous genes are used.

The gfp gene has been integrated into the genome by single cross-over integration. Therefore the possibility exists of reversion to the wild-type genotype by removal of the integrated DNA construct.
Additional remarks selection procedure
Other details transgene
Promoter
Gene Model of Parasite PBANKA_0416100
Gene Model P. falciparum ortholog PF3D7_0905300
Gene productdynein heavy chain, putative
Gene product: Alternative name
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 1aaGATATCatatttaaacagattaagtaccg
Additional information primer 1forward
Sequence Primer 2aaGGATCCtttttatcatttggataattaattc
Additional information primer 2reverse
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 / InsertionInsertion locus
Gene Model of Parasite Not available
Gene productNot available
Gene product: Alternative namesmall subunit ribosomal rna gene (c- or d-type unit)
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