Summary

RMgm-131
Malaria parasiteP. berghei
Genotype
Transgene
Transgene not Plasmodium: GFP (gfp-mut3)
Promoter: Gene model: PBANKA_0925400; Gene model (P.falciparum): PF3D7_1122900; 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, 13:53
  *RMgm-131
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-131
Principal name500
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 PB000733.03.0 (PF11_0240; dynein heavy chain, putative)

Protein (function)
PB000733.03.0 (PF11_0240; 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 prescence 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 PB000733.03.0 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 1576 bp fragment (HpaI/BamHI) of the promoter region of PB000733.03.0.
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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AACACACGTAAATGTTTTAATGCACACATATATGTATGAATATGTATGCTTAGAAGAGAG
ACTCACATACATGCGATTCAAATTCGTGTGATTCGTCGAGAATTCTATTTCGTTTTGTTA
TTTAACTAAATTTGGACAGTAAAAAATATGCTCTAAACTATTATGTTTTATCCTTGTAGG
ACGAAGACGGTTATATTAGTAAAAGTGATCTTGCCACATTAGTTCATGACAATGGTGTTA
ATAATAAAGATATAGAGAATTTTTTCATTTCTGTGCATAGCATTAAAAAAAACATAACTA
AGGACAAGAAGATAAATAAAGTAATAATTTATTCCCTATTTTAAATAAAATTATTTTAGT
TCGAATCCTATATTTTATATAATATATTTTTTATTTATGCAGATAAGTTTTGAAGATTTT
AAGGATTACATGTTGAGCACATTTTGAACCCATATAAAAATTATATTCCTAAATAATAAT
ATAGATAAAATACCGCCAAGAAAATTAATAGTTAAAAAGAAAAAGTATATGAATGCGTTT
TCTTATGAATTAAGGGATAAAGGTGAATAATTATTTTTCTTTATATATTTTTTTATTGTA
TACCACTTTAAAACGAAAGAATTTATAGTTTAAAATTTAATTAACAAAATATTTGTATGT
TATTTAACATTTTATAATTCGTTTATCTTAATGCCTTATCATTTTATTACCCCATCCTCT
CTCCTTTATATATATTATTATTTTCCTATAATCTTTATATTTATTTATTACATTCATTTT
TGAAAATTGCATAAAAATATCTCAAATTAATGTATATTTTATTATTATTGATACATTAAT
TATTTTTTTTCAAGTCTTTTTTTCATTACATTTTTTATATAAATTTATTATGATGAATTA
ATGTTAAAATGGAAGGAAAATAATTTTAATGTGTAGATGCATACATATATGTATTCTCAT
GTAAAACATTTTCTATATGATGCTTTAATAGATGCATCTTTTTATTTTTTTGTACGTAGT
TTATATTTTGCATTGAAACTATTAAAATTCACCCCCAATAATAAAACGTTTAAAAACTAT
AAAGCACATCGCTATATTGTTATAATTTTACTAATTCTATATAATATCATTAAATTTTAA
CACACGTAAAAATAGACAAAATATATACTAGATATCATATGCATGTTTAAAATCCACGCA
ATTAGTTAGTATGTAAATATATGCTTCAATTTACGTACATCTTGCACATTTAAAATTAGT
GAAAAAAAAAACAAAATAAAAATATATTATATGGCCTTTTTTCCTTAACACAAAATATTT
CATAAACGCATGTATAAGTGTTTTTTTCTTTTTTTTAATCTTTAAATAATTTCATAAAGA
TAAGCTAAAAAAAAATAAATAGTAAACGAAGTGATGCTACTAGTAATAATAAGTGTCTAG
AAGTACAGCATCATAATTGCAGAATTAGTAATAACACTAGAATATATAAAAATTAATAGT
AATAAAAGTAGTAATAGTAGCAAAATCGTGTATCCATTGTCACTTTCTAAATATGTGTAT
ATATAGGTTTGTATAAAATGGATCCATGAGTAAAGGAGAAGAACTTTTCACTGGAGTTGT
CCCAATTCTTGTTGAATTAGATGGTGATGTTAATGGGCACAAATTTTCTGTCAGTGGAGA
GGGTGAAGGTGATGCAACATACGGAAAACTTACCCTTAAATTTATTTGCACTACTGGAAA
ACTACCTGTTCCATGGCCAACACTTGTCACTACTTTCGGTTATGGTGTTCAATGCTTTGC
GAGATACCCAGATCATATGAAACAGCATGACTTTTTCAAGAGTGCCATGCCCGAAGGTTA
TGTACAGGAAAGAACTATATTTTTCAAAGATGACGGGAACTACAAGACACGTGCTGAAGT
CAAGTTTGAAGGTGATACCCTTGTTAATAGAATCGAGTTAAAAGGTATTGATTTTAAAGA
AGATGGAAACATTCTTGGACACAAATTGGAATACAACTATAACTCACACAATGTATACAT
CATGGCAGACAAACAAAAGAATGGAATCAAAGTTAACTTCAAAATTAGACACAACATTGA
AGATGGAAGCGTTCAACTAGCAGACCATTATCAACAAAATACTCCAATTGGCGATGGCCC
TGTCCTTTTACCAGACAACCATTACCTGTCCACACAATCTGCCCTTTCGAAAGATCCCAA
CGAAAAGAGAGACCACATGGTCCTTCTTGAGTTTGTAACAGCTGCTGGGATTACACATGG
CATGGATGAACTATACAAATAAATTCTAGAAGATCCCGTTTTTCTTACTTATATATTTAT
ACCAATTGATTGTATTTATAACTGTAAAAATGTGTATGTTGTGTGCATATTTTTTTTTGT
GCATGCACATGCATGTAAATAGCTAAAATTATGAACATTTTATTTTTTGTTCAGAAAAAA
AAAACTTTACACACATAAAATGGCTAGTATGAATAGCCATATTTTATATAAATTAAATCC
TATGAATTTATGACCATATTAAAAATTTAGATATTTATGGAACATAATATGTTTGAAACA
ATAAGACAAAATTATTATTATTATTATTATTTTTACTGTTATAATTATGTTGTCTCTTCA
ATGATTCATAAATAGTTGGACTTGATTTTTAAAATGTTTATAATATGATTAGCATAGTTA
AATAAAAAAAGTTGAAAAATTAAAAAAAAACATATAAACACAAATGATGTTTTTTCCTTC
AATTTCGGGTACCGAGCTCGAATTCAACCTGGTTGATCTTGCCAGTAGTCATATGCTTGT
CTCAAAGATTAAGCCATGCAAGTGAAAGTATATGCACATTTATTGCAGAAACTGCGAACG
GCTCATTAAAACAGTTATAATCTACTTGACATTTTATTATAAGGATAACTACGGAAAAGC
TGTAGCTAATACTTGTTAAGTACTTTTACTCCCCGGAGTAATTGTATGTATTTGTTAAGA
CCCCTAAGAAAAAATGATATTAAAGGAATTATAACAAAGAAGCAACACATAATATAATTA
TTCAGTGTGTATCAATCGAGTTTCTGACCTATCAGCTTTTGATGTTAGGGTATTGACCTA
ACATGGCTTTGACGGGTAACGGGGAATTAGAGTTCGATTCCGGAGAGGGAGCCTGAGAAA
TAGCTACCACATCTAAGGAAGGCAGCAGGCGCGTAAATTACCCAATTCTAAATAAGAGAG
GTAGTGACAAGAAATAACAATATAAGGCCAAATTTTGGTTTTATAATTGGAATGATGGGA
ATTTAAAACCTTCCCAAAAATCAATTGGAGGGCAAGTCTGGTGCCAGCAGCCGCGGTAAT
TCCAGCTCCAATAGCGTATATTAAAATTGTTGCAGTTAAAACGCTCGTAGTTGAACTTCA
AGGGTATAATTATTTTAAGCAACTCACTTGGAAAGAATCATGACTTCTGTCACTGCTTTT
ATCCTTGTTGCAGTTCTTTTAATACAGGGCCCTTTGAGAGCCCATTAATTTATGACTGGG
TTTCTCGTTACTTTGAGTAAATTAGAGTGTTTAAAGCAAACAGATAAAGCGTATTTTACT
GTGTTTGAATACTATAGCATGGAATAACAACATTGAATAGGTCAAAAGTTTTTGAAAAAT
TTTTCTTATTTTGGCTTAGATACAGTTAATAGGAGTAGCTTGGGGGCATTTGTATTCAGA
TGTCAGAGGTGAAATTCTTAGATTTTCTGGAGACAAACAACTGCGAAAGCATTTGCCTAA
AATACTTCCATTAATCAAGAACGAAAGTTAAGGGAGTGAAGACGATCAGATACCGTCGTA
ATCTTAACCATAAACTATGCCGACTAAGTGTTGGATGAAAATTTATAAATAAAACTATCT
TCTTTAAAGGAGTAGTTTTTTAGATGCTTCCTTCAGTACCTTATGAGAAATCAAAGTCTT
TGGGTTCTGGGGCGAGTATTCGCGCAAGCGAGAAAGTTAAAAGAATTGACGGAAGGGCAC
CACCAGGCGTGGAGCTTGCGGCTTAATTTGACTCAACACGGGGAAACTCACTAGTTTAAG
ACAAGAGTAGGATTGACAGATTAATAGCTCTTTCTTGATTTCTTGGATGGTGATGCATGG
CCGTTTTTAGTTCGTGAATATGATTTGTCTGGTTAATTCCGATAACGAACGAGATCTTAA
CCTGCTAATTAGCGGTGGATATGTGATATTCTTCGAAGGTGGACTAACTATAGCGTTTTC
GAAGGTATGTTGCATAATCAAATTGGTTTACCCTTTGTTTTTTTGTAGCATATTCTTTTA
TTTCGTTGGGTTTTTTCCCTAGTAAGGATGTATCTGCTTTATTTAATGCTTCTTAGAGGA
ACGATGTGTGTCTAACACAAGGAAGTTTAAGGCAACAACAGGTCTGTGATGTCCTTAGAT
ATACTAGGCTGCACGCGTGCTACACTGATATGTAAAACGAGTATTTAAAATTATATCTGT
ATGGTAGATAATTTAATTTCTACGTATTATCAGCATATACTTTTCCTACACTGAAATAGT
GAAGGTAATCTTTATCAATACATATCGTGATGGGGATAGATTATTGCAATTATTAATCTT
GAACGAGGAATGCCTAGTAAGCATGATTCATCAGATTGTGCTGACTACGTCCCTGCCCTT
TGTACACACCGCCCGTCGCTCCTACCGATTGAAAGATATGATGAATTGTTTGGACAAGAA
AATAGAAATTTTATTTTTATTTTTTTTGGAAGGACCGTAAATCCTATCTTTTAAAGGAAG
GAGAAGTCGTAACAAGGTTTCCGTAGGTGAATTCACTGGCCGTCGTTTTACAACGTCGTG
ACTGGGAAAACCCTGGCGTTACCCAACTTAATCGCCTTGCAGCACATCCCCCTTTCGCCA
GCTGGCGTAATAGCGAAGAGGCCCGCACCGATCGCCCTTCCCAACAGTTGCGCAGCCTGA
ATGGCGAATGGCGCCTGATGCGGTATTTTCTCCTTACGCATCTGTGCGGTATTTCACACC
GCATATGGTGCACTCTCAGTACAATCTGCTCTGATGCCGCATAGTTAAGCCAGCCCCGAC
ACCCGCCAACACCCGCTGACGCGCCCTGACGGGCTTGTCTGCTCCCGGCATCCGCTTACA
GACAAGCTGTGACCGTCTCCGGGAGCTGCATGTGTCAGAGGTTTTCACCGTCATCACCGA
AACGCGCGAGACGAAAGGGCCTCGTGATACGCCTATTTTTATAGGTTAATGTCATGATAA
TAATGGTTTCTTAGACGTCAGGTGGCACTTTTCGGGGAAATGTGCGCGGAACCCCTATTT
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
TACAGCGTGAGCATTGAGAAAGCGCCACGCTTCCCGAAGGGAGAAAGGCGGACAGGTATC
CGGTAAGCGGCAGGGTCGGAACAGGAGAGCGCACGAGGGAGCTTCCAGGGGGAAACGCCT
GGTATCTTTATAGTCCTGTCGGGTTTCGCCACCTCTGACTTGAGCGTCGATTTTTGTGAT
GCTCGTCAGGGGGGCGGAGCCTATGGAAAAACGCCAGCAACGCGGCCTTTTTACGGTTCC
TGGCCTTTTGCTGGCCTTTTGCTCACATGTTCTTTCCTGCGTTATCCCCTGATTCTGTGG
ATAACCGTATTACCGCCTTTGAGTGAGCTGATACCGCTCGCCGCAGCCGAACGACCGAGC
GCAGCGAGTCAGTGAGCGAGGAAGCGGAAGAGCGCCCAATACGCAAACCGCCTCTCCCCG
CGCGTTGGCCGATTCATTAATGCAGCTGGCACGACAGGTTTCCCGACTGGAAAGCGGGCA
GTGAGCGCAACGCAATTAATGTGAGTTAGCTCACTCATTAGGCACCCCAGGCTTTACACT
TTATGCTTCCGGCTCGTATGTTGTGTGGAATTGTGAGCGGATAACAATTTCACACAGGAA
ACAGCTATGACCATGATTACGCCAAGCTTGCATGCCTGCAGGTCAACAATAAATAATAAA
TAAATATTGTGGAAATAAAATAACATATAATTATTTTTAATACATTGATTTCCCTTTTAT
TTTTTTAAATTTCATTGATATAAAAATATATAATAATAACATATATGATTTCAAATTAAT
CTTTTCAAAAATGGTGTTTATTTTTGTATGTTTGTGTATGAATTAATCACATAACACATC
TATTAAATTGAGTTGGTAATATAGACACAAATAAATATATATATTTTTATAGCTTAAAAG
TGTGTTATGAATATTTTAAGCATATTTTCTTTTTCTTTGGATTGTGTAAAATGAACTCAT
ATAATGCGTTTTTTTGTTTTTGTTATTTTGTCATTTTGTTATTTTGCTATTTTATGGATT
AATTTTTGTTTATAAAATGGGAAATAATTTTAACATATTTAAATAAATGGAGAAAAAATA
TAAAATAATTATAAAAAAAAGTTAATACACATTTTTTCCTGTTATAGACCTTATATTTAT
TTATCCATATATATATATATATATATATATATATATATATACATACCAAGTGAATTAAGA
GGAAAGCTAATTTATTATTCAGAATAATATATGAACTATATATAATTTTTATTATTTTGG
TGTATATTAATCTGTCTATATGCATACATGCAATAATTTATCGACTTATATATCAAATAA
CATAAAATAGAAGTGTTTTAAATTATGGATATATGCTCAATATTCATTTTTTTTAATAAG
TTAGCTATATTTAAATTATACATTTTATATATGGTCTCTTTTTTTTTTAAATATTATTTA
AGTGATCATGAAAATATAAATAATTTTTTTTTATTTAATATCCTTTTGCTTGCATGTGGT
AAATGGAAATTTGGATGTGTTTTGAARGTTCGGATATAGTTGTATGGACATATAATATAT
TTTGTGAAAAATTGGTTTTATGTTTATACTTATGCCAATACTTTTTGAGTAAAACAAAGC
AAGTGCTTATAAATAATTAAAGCCAATTTTATAATATATATTTTTTTATTTAATTTGAAT
TTAGTAGTATAATTTTTTATGGTAAGTGCTCAAAGAGAGTTGCTTATAAAGTATGGTTTG
TTTCTTTTTCGCCATTTTGAATTACACATTAAAAATATATAGATACATATATTATAATAT
GAAATCATTAATAATTTAGGGAAATTCTACAAATTTAAAAACGAATAAAATAATTGTTTT
TCATCATGCCATAACACAATATTGATATATACATGTACAAACATTTTTTTTATTTGGAAA
ATATAAATTATATAAAAAAAAATGTATAGTATACAAAATGAGCATATTCACACGGGGTGG
ACGTTCATTTTTTCATTTTTCCCCTGTTTTTTATGAGTATATGATAAAATTTTATGAACA
TTTACACAAAATGAAAATGGATATATAGGAAAAATGGAGCGGTATTTCATTTATCTTTGA
TTGTCATTTGGATATTATATTACCYTGGGTAGGCAATTAAAAATGTTAAATAACAATTTA
AGGAAATTATATTTTATATATTAAAATTAACACTGTATTATATGATTCGCTTATAAAAGC
CACTCTTTCCCCATGCAAAGCTGTTTAATATCAATTTTAACAAATTACACACATGTTAAT
ATATTTATATATATAATTTATATATTTATAATTTATATATTTATATTTTTATTATTTATA
TATTTATTATTTATTGTGTGTGTCAATTCGGGTAGGATATACCTCTTTTTTATTGTTTAA
AGCGATTTGTATTCTAAAATATAAAGRATTTGAAAAAGAGAAAGATAGAATATGATCCCA
TCATATATAGCCCTATAATTTTTATTTAGCAGCGAATTAATTTTTCTATTAAGTTTATGT
GTAATTAAAATAACGGAATATATATAATACAATAAAAAAGTGCATAAATTAAAATTTTTT
CAATTAAATTTTTTTTTTTAAGGGGTTATATAATATTAAATATATAAAATACGATTATAT
ATTTTTGCTACAATTTTTTATATTAAGATATAAATAGTAAATAAATGGTATTATATGGCA
TGTAATATATAAATTTTTTCCAATTTTTATTTTATATACACTTTTCCTTTTTTTGTCATA
AAACTTAAACAATTTACACATTCATTTTAAAAATTGACTATTTGTTTCAACATTTTTTGA
GTTTCCGTTTTATAATAGTATTTTCATTTGTATATTGCTTATATATATAAATACACACCT
AAATGTTACAAAGGATCAATGCATAAACCGGTGTGTCTGGTCGTCGCGATGACCCCCAAG
AGGGGCATCGGCATCAACAACGGCCTCCCGTGGCCCCACTTGACCACAGATTTCAAACAC
TTTTCTCGTGTGACAAAAACGACGCCCGAAGAAGCCAGTCGCCTGAACGGGTGGCTTCCC
AGGAAATTTGCAAAGACGGGCGACTCTGGACTTCCCTCTCCATCAGTCGGCAAGAGATTC
AACGCCGTTGTCATGGGACGGAAAACCTGGGAAAGCATGCCTCGAAAGTTTAGACCCCTC
GTGGACAGATTGAACATCGTCGTTTCCTCTTCCCTCAAAGAAGAAGACATTGCGGCGGAG
AAGCCTCAAGCTGAAGGCCAGCAGCGCGTCCGAGTCTGTGCTTCACTCCCAGCAGCTCTC
AGCCTTCTGGAGGAAGAGTACAAGGATTCTGTCGACCAGATTTTTGTCGTGGGAGGAGCG
GGACTGTACGAGGCAGCGCTGTCTCTGGGCGTTGCCTCTCACCTGTACATCACGCGTGTA
GCCCGCGAGTTTCCGTGCGACGTTTTCTTCCCTGCGTTCCCCGGAGATGACATTCTTTCA
AACAAATCAACTGCTGCGCAGGCTGCAGCTCCTGCCGAGTCTGTGTTCGTTCCCTTTTGT
CCGGAGCTCGGAAGAGAGAAGGACAATGAAGCGACGTATCGACCCATCTTCATTTCCAAG
ACCTTCTCAGACAACGGGGTTCCCTACGACTTTGTGGTTCTCGAGAAGAGAAGGAAGACT
GACGACGCAGCCACTGCGGAACCGAGCAACGCAATGAGCTCCTTGACGTCCACGAGGGAG
ACAACTCCCGTGCACGGGTTGCAGGCTCCTTCTTCGGCCGCAGCCATTGCCCCGGTGTTG
GCGTGGATGGACGAAGAAGACCGGAAAAAACGCGAGCAAAAGGAACTGATTCGGGCCGTT
CCGCATGTTCACTTTAGAGGCCATGAAGAGTTCCAGTACCTTGATCTCATTGCCGACATT
ATTAACAATGGAAGGACAATGGATGACCGAACGGGCGTTGGTGTCATCTCCAAATTCGGC
TGCACTATGCGCTACTCGCTGGATCAGGCCTTTCCACTTCTCACCACAAAGCGTGTGTTC
TGGAAAGGGGTCCTCGAAGAGTTGCTGTGGTTCATTCGCGGCGACACGAACGCAAACCAT
CTTTCTGAGAAGGGCGTGAAGATCTGGGACAAGAATGTGACACGCGAGTTCCTCGATTCG
CGCAATCTCCCCCACCGAGAGGTCGGAGACATCGGCCCGGGCTACGGCTTCCAGTGGAGA
CACTTCGGCGCGGCATACAAAGACATGCACACAGACTACACAGGGCAGGGCGTCGACCAG
CTGAAGAATGTGATCCAGATGCTGAGAACGAATCCAACAGATCGTCGCATGCTCATGACT
GCCTGGAATCCTGCAGCGCTGGACGAAATGGCGCTGCCGCCTTGTCACTTGTTGTGCCAG
TTCTACGTGAACGACCAGAAGGAGCTGTCGTGCATCATGTATCAGCGGTCGTGCGATGTC
GGCCTCGGCGTCCCCTTCAACATCGCTTCCTATTCGCTTTTGACGCTCATGGTTGCACAC
GTCTGCAACCTAAAACCTAAGGAGTTCATTCACTTCATGGGGAACACGCATGTCTACACG
AACCATGTCGAGGCTTTAAAAGAGCAGCTGCGGAGAGAACCGAGACCGTTCCCCATTGTG
AACATCCTCAACAAGGAACGCATCAAGGAAATCGACGATTTCACCGCCGAGGATTTTGAG
GTCGTGGGCTACGTCCCGCACGGACGAATCCAGATGGAGATGGCTGTCTAGCGGAAATAC
AGAAGCTAGCTTTGATCCCGTTTTTCTTACTTATATATTTATACCAATTGATTGTATTTA
TAACTGTAAAAATGTGTATGTTGTGTGCATATTTTTTTTTGTGCATGCACATGCATGTAA
ATAGCTAAAATTATGAACATTTTATTTTTTGTTCAGAAAAAAAAAACTTTACACACATAA
AATGGCTAGTATGAATAGCCATATTTTATATAAATTAAATCCTATGAATTTATGACCATA
TTAAAAATTTAGATATTTATGGAACATAATATGTTTGAAACAATAAGACAAAATTATTAT
TATTATTATTATTTTTACTGTTATAATTATGTTGTCTCTTCAATGATTCATAAATAGTTG
GACTTGATTTTTAAAATGTTTATAATATGATTAGCATAGTTAAATAAAAAAAGTTGAAAA
ATTAAAAAAAAACATATAAACACAAATGATGTTTTTTCCTTCAATTTCGAT
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 PB000733.03.0 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 1576 bp fragment (HpaI/BamHI) of the promoter region of PB000733.03.0.

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_0925400
Gene Model P. falciparum ortholog PF3D7_1122900
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 1aaGTTAACacacgtaaatgttttaatgcac
Additional information primer 1forward
Sequence Primer 2aaGGATCCattttatacaaacctatatatacac
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