Summary

RMgm-132
Malaria parasiteP. berghei
Genotype
Transgene
Transgene not Plasmodium: GFP (gfp-mut3)
Promoter: Gene model: PBANKA_0619200; Gene model (P.falciparum): PF3D7_0721700; Gene product: secreted ookinete protein, putative (PSOP1, Putative secreted ookinete protein 1)
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, 14:31
  *RMgm-132
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-132
Principal name479
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 and female gametocytes. No expression could be detected in asexual blood stages.
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 PB000198.00.0 (PF07_0089, hypothetical protein conserved; PSOP1)

Protein (function)
PB000198.00.0 (PF07_0089, hypothetical protein conserved; PSOP1). The protein is detected in a proteome analysis of ookinetes and contains a predicted signal sequence. See also RMgm-88 for a mutant lacking expression of PB000198.00.0 (PSOP1).

Phenotype
Phenotype analyses indicate absence of promoter activity in asexual blood stages and active transcription in both male and female gametocytes. The absence in asexual blood stages and presence in males and females 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 PB000198.00.0 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 754 bp fragment (SmaI/ BamHI) of the promoter region of PB000198.00.0.
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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GGGCAATGTGATAAAATTAAGATTGGGATATAAGACAATCTATGAATTAATTTTTGTAAT
TTTCTTTTTATTTTGAAGAATTTTTTGCTTTTTGTCTTAAAAATGCAATAATTGTTTATA
TAAAAAAAATATTCACGTGTTAATAAGTATTTGTCTATAATATGAATTTTATTAATTTTT
TTATGACTAAAACATAATTATTTCGATTTTTCAATTATTTTTGGCTAAATATACAAAAAA
GGCGGAAAAGAGTTAGAAAAAAAGTTAGAAAAAAGGTTAGAAAAAAACATATACATGTAT
GTACACCTGAACAGCTCATAATTTTGGCTATTTTTTTTTTCCAAAAGTAGCTAATTGCTC
ATTTTGGGGGGCTATATAGCGAATATTTCACTGTATACTAGCTTTTTTCCCTTTTTATTT
ATAATTTAAATTTTTATTAAAATATATTTTTTGTTATAATTATAATTTTATTATATATAT
ATTTTTTTTTATTTATATTTTTATATTTATTTTTATATCTTTATGTTTATATTTTTATAT
TTCTTTATTTTGATCATTTTTCAAACTTGGGTGTTTTAATATTACCAATTCGATTTAGCT
TTGTGTGCGCAATTTTTACTATATTATATTTTTTTGTCTTCTCAAGTACTCAAACATACG
CTATTTAAATTTACAAAACAAGACATCGTCCAGACACACACTATACATACATATACAAAA
TCGTTAACATCTCGAATCTTAATAAGACAGATATAAAAGAATCCTTAAGGGCGAATTCCA
GCACACTGGCGGCCGTTACTAGTGGATCCATGAGTAAAGGAGAAGAACTTTTCACTGGAG
TTGTCCCAATTCTTGTTGAATTAGATGGTGATGTTAATGGGCACAAATTTTCTGTCAGTG
GAGAGGGTGAAGGTGATGCAACATACGGAAAACTTACCCTTAAATTTATTTGCACTACTG
GAAAACTACCTGTTCCATGGCCAACACTTGTCACTACTTTCGGTTATGGTGTTCAATGCT
TTGCGAGATACCCAGATCATATGAAACAGCATGACTTTTTCAAGAGTGCCATGCCCGAAG
GTTATGTACAGGAAAGAACTATATTTTTCAAAGATGACGGGAACTACAAGACACGTGCTG
AAGTCAAGTTTGAAGGTGATACCCTTGTTAATAGAATCGAGTTAAAAGGTATTGATTTTA
AAGAAGATGGAAACATTCTTGGACACAAATTGGAATACAACTATAACTCACACAATGTAT
ACATCATGGCAGACAAACAAAAGAATGGAATCAAAGTTAACTTCAAAATTAGACACAACA
TTGAAGATGGAAGCGTTCAACTAGCAGACCATTATCAACAAAATACTCCAATTGGCGATG
GCCCTGTCCTTTTACCAGACAACCATTACCTGTCCACACAATCTGCCCTTTCGAAAGATC
CCAACGAAAAGAGAGACCACATGGTCCTTCTTGAGTTTGTAACAGCTGCTGGGATTACAC
ATGGCATGGATGAACTATACAAATAAATTCTAGAAGATCCCGTTTTTCTTACTTATATAT
TTATACCAATTGATTGTATTTATAACTGTAAAAATGTGTATGTTGTGTGCATATTTTTTT
TTGTGCATGCACATGCATGTAAATAGCTAAAATTATGAACATTTTATTTTTTGTTCAGAA
AAAAAAAACTTTACACACATAAAATGGCTAGTATGAATAGCCATATTTTATATAAATTAA
ATCCTATGAATTTATGACCATATTAAAAATTTAGATATTTATGGAACATAATATGTTTGA
AACAATAAGACAAAATTATTATTATTATTATTATTTTTACTGTTATAATTATGTTGTCTC
TTCAATGATTCATAAATAGTTGGACTTGATTTTTAAAATGTTTATAATATGATTAGCATA
GTTAAATAAAAAAAGTTGAAAAATTAAAAAAAAACATATAAACACAAATGATGTTTTTTC
CTTCAATTTCGGGTACCGAGCTCGAATTCAACCTGGTTGATCTTGCCAGTAGTCATATGC
TTGTCTCAAAGATTAAGCCATGCAAGTGAAAGTATATGCACATTTATTGCAGAAACTGCG
AACGGCTCATTAAAACAGTTATAATCTACTTGACATTTTATTATAAGGATAACTACGGAA
AAGCTGTAGCTAATACTTGTTAAGTACTTTTACTCCCCGGAGTAATTGTATGTATTTGTT
AAGACCCCTAAGAAAAAATGATATTAAAGGAATTATAACAAAGAAGCAACACATAATATA
ATTATTCAGTGTGTATCAATCGAGTTTCTGACCTATCAGCTTTTGATGTTAGGGTATTGA
CCTAACATGGCTTTGACGGGTAACGGGGAATTAGAGTTCGATTCCGGAGAGGGAGCCTGA
GAAATAGCTACCACATCTAAGGAAGGCAGCAGGCGCGTAAATTACCCAATTCTAAATAAG
AGAGGTAGTGACAAGAAATAACAATATAAGGCCAAATTTTGGTTTTATAATTGGAATGAT
GGGAATTTAAAACCTTCCCAAAAATCAATTGGAGGGCAAGTCTGGTGCCAGCAGCCGCGG
TAATTCCAGCTCCAATAGCGTATATTAAAATTGTTGCAGTTAAAACGCTCGTAGTTGAAC
TTCAAGGGTATAATTATTTTAAGCAACTCACTTGGAAAGAATCATGACTTCTGTCACTGC
TTTTATCCTTGTTGCAGTTCTTTTAATACAGGGCCCTTTGAGAGCCCATTAATTTATGAC
TGGGTTTCTCGTTACTTTGAGTAAATTAGAGTGTTTAAAGCAAACAGATAAAGCGTATTT
TACTGTGTTTGAATACTATAGCATGGAATAACAACATTGAATAGGTCAAAAGTTTTTGAA
AAATTTTTCTTATTTTGGCTTAGATACAGTTAATAGGAGTAGCTTGGGGGCATTTGTATT
CAGATGTCAGAGGTGAAATTCTTAGATTTTCTGGAGACAAACAACTGCGAAAGCATTTGC
CTAAAATACTTCCATTAATCAAGAACGAAAGTTAAGGGAGTGAAGACGATCAGATACCGT
CGTAATCTTAACCATAAACTATGCCGACTAAGTGTTGGATGAAAATTTATAAATAAAACT
ATCTTCTTTAAAGGAGTAGTTTTTTAGATGCTTCCTTCAGTACCTTATGAGAAATCAAAG
TCTTTGGGTTCTGGGGCGAGTATTCGCGCAAGCGAGAAAGTTAAAAGAATTGACGGAAGG
GCACCACCAGGCGTGGAGCTTGCGGCTTAATTTGACTCAACACGGGGAAACTCACTAGTT
TAAGACAAGAGTAGGATTGACAGATTAATAGCTCTTTCTTGATTTCTTGGATGGTGATGC
ATGGCCGTTTTTAGTTCGTGAATATGATTTGTCTGGTTAATTCCGATAACGAACGAGATC
TTAACCTGCTAATTAGCGGTGGATATGTGATATTCTTCGAAGGTGGACTAACTATAGCGT
TTTCGAAGGTATGTTGCATAATCAAATTGGTTTACCCTTTGTTTTTTTGTAGCATATTCT
TTTATTTCGTTGGGTTTTTTCCCTAGTAAGGATGTATCTGCTTTATTTAATGCTTCTTAG
AGGAACGATGTGTGTCTAACACAAGGAAGTTTAAGGCAACAACAGGTCTGTGATGTCCTT
AGATATACTAGGCTGCACGCGTGCTACACTGATATGTAAAACGAGTATTTAAAATTATAT
CTGTATGGTAGATAATTTAATTTCTACGTATTATCAGCATATACTTTTCCTACACTGAAA
TAGTGAAGGTAATCTTTATCAATACATATCGTGATGGGGATAGATTATTGCAATTATTAA
TCTTGAACGAGGAATGCCTAGTAAGCATGATTCATCAGATTGTGCTGACTACGTCCCTGC
CCTTTGTACACACCGCCCGTCGCTCCTACCGATTGAAAGATATGATGAATTGTTTGGACA
AGAAAATAGAAATTTTATTTTTATTTTTTTTGGAAGGACCGTAAATCCTATCTTTTAAAG
GAAGGAGAAGTCGTAACAAGGTTTCCGTAGGTGAATTCACTGGCCGTCGTTTTACAACGT
CGTGACTGGGAAAACCCTGGCGTTACCCAACTTAATCGCCTTGCAGCACATCCCCCTTTC
GCCAGCTGGCGTAATAGCGAAGAGGCCCGCACCGATCGCCCTTCCCAACAGTTGCGCAGC
CTGAATGGCGAATGGCGCCTGATGCGGTATTTTCTCCTTACGCATCTGTGCGGTATTTCA
CACCGCATATGGTGCACTCTCAGTACAATCTGCTCTGATGCCGCATAGTTAAGCCAGCCC
CGACACCCGCCAACACCCGCTGACGCGCCCTGACGGGCTTGTCTGCTCCCGGCATCCGCT
TACAGACAAGCTGTGACCGTCTCCGGGAGCTGCATGTGTCAGAGGTTTTCACCGTCATCA
CCGAAACGCGCGAGACGAAAGGGCCTCGTGATACGCCTATTTTTATAGGTTAATGTCATG
ATAATAATGGTTTCTTAGACGTCAGGTGGCACTTTTCGGGGAAATGTGCGCGGAACCCCT
ATTTGTTTATTTTTCTAAATACATTCAAATATGTATCCGCTCATGAGACAATAACCCTGA
TAAATGCTTCAATAATATTGAAAAAGGAAGAGTATGAGTATTCAACATTTCCGTGTCGCC
CTTATTCCCTTTTTTGCGGCATTTTGCCTTCCTGTTTTTGCTCACCCAGAAACGCTGGTG
AAAGTAAAAGATGCTGAAGATCAGTTGGGTGCACGAGTGGGTTACATCGAACTGGATCTC
AACAGCGGTAAGATCCTTGAGAGTTTTCGCCCCGAAGAACGTTTTCCAATGATGAGCACT
TTTAAAGTTCTGCTATGTGGCGCGGTATTATCCCGTATTGACGCCGGGCAAGAGCAACTC
GGTCGCCGCATACACTATTCTCAGAATGACTTGGTTGAGTACTCACCAGTCACAGAAAAG
CATCTTACGGATGGCATGACAGTAAGAGAATTATGCAGTGCTGCCATAACCATGAGTGAT
AACACTGCGGCCAACTTACTTCTGACAACGATCGGAGGACCGAAGGAGCTAACCGCTTTT
TTGCACAACATGGGGGATCATGTAACTCGCCTTGATCGTTGGGAACCGGAGCTGAATGAA
GCCATACCAAACGACGAGCGTGACACCACGATGCCTGTAGCAATGGCAACAACGTTGCGC
AAACTATTAACTGGCGAACTACTTACTCTAGCTTCCCGGCAACAATTAATAGACTGGATG
GAGGCGGATAAAGTTGCAGGACCACTTCTGCGCTCGGCCCTTCCGGCTGGCTGGTTTATT
GCTGATAAATCTGGAGCCGGTGAGCGTGGGTCTCGCGGTATCATTGCAGCACTGGGGCCA
GATGGTAAGCCCTCCCGTATCGTAGTTATCTACACGACGGGGAGTCAGGCAACTATGGAT
GAACGAAATAGACAGATCGCTGAGATAGGTGCCTCACTGATTAAGCATTGGTAACTGTCA
GACCAAGTTTACTCATATATACTTTAGATTGATTTAAAACTTCATTTTTAATTTAAAAGG
ATCTAGGTGAAGATCCTTTTTGATAATCTCATGACCAAAATCCCTTAACGTGAGTTTTCG
TTCCACTGAGCGTCAGACCCCGTAGAAAAGATCAAAGGATCTTCTTGAGATCCTTTTTTT
CTGCGCGTAATCTGCTGCTTGCAAACAAAAAAACCACCGCTACCAGCGGTGGTTTGTTTG
CCGGATCAAGAGCTACCAACTCTTTTTCCGAAGGTAACTGGCTTCAGCAGAGCGCAGATA
CCAAATACTGTCCTTCTAGTGTAGCCGTAGTTAGGCCACCACTTCAAGAACTCTGTAGCA
CCGCCTACATACCTCGCTCTGCTAATCCTGTTACCAGTGGCTGCTGCCAGTGGCGATAAG
TCGTGTCTTACCGGGTTGGACTCAAGACGATAGTTACCGGATAAGGCGCAGCGGTCGGGC
TGAACGGGGGGTTCGTGCACACAGCCCAGCTTGGAGCGAACGACCTACACCGAACTGAGA
TACCTACAGCGTGAGCATTGAGAAAGCGCCACGCTTCCCGAAGGGAGAAAGGCGGACAGG
TATCCGGTAAGCGGCAGGGTCGGAACAGGAGAGCGCACGAGGGAGCTTCCAGGGGGAAAC
GCCTGGTATCTTTATAGTCCTGTCGGGTTTCGCCACCTCTGACTTGAGCGTCGATTTTTG
TGATGCTCGTCAGGGGGGCGGAGCCTATGGAAAAACGCCAGCAACGCGGCCTTTTTACGG
TTCCTGGCCTTTTGCTGGCCTTTTGCTCACATGTTCTTTCCTGCGTTATCCCCTGATTCT
GTGGATAACCGTATTACCGCCTTTGAGTGAGCTGATACCGCTCGCCGCAGCCGAACGACC
GAGCGCAGCGAGTCAGTGAGCGAGGAAGCGGAAGAGCGCCCAATACGCAAACCGCCTCTC
CCCGCGCGTTGGCCGATTCATTAATGCAGCTGGCACGACAGGTTTCCCGACTGGAAAGCG
GGCAGTGAGCGCAACGCAATTAATGTGAGTTAGCTCACTCATTAGGCACCCCAGGCTTTA
CACTTTATGCTTCCGGCTCGTATGTTGTGTGGAATTGTGAGCGGATAACAATTTCACACA
GGAAACAGCTATGACCATGATTACGCCAAGCTTGCATGCCTGCAGGTCAACAATAAATAA
TAAATAAATATTGTGGAAATAAAATAACATATAATTATTTTTAATACATTGATTTCCCTT
TTATTTTTTTAAATTTCATTGATATAAAAATATATAATAATAACATATATGATTTCAAAT
TAATCTTTTCAAAAATGGTGTTTATTTTTGTATGTTTGTGTATGAATTAATCACATAACA
CATCTATTAAATTGAGTTGGTAATATAGACACAAATAAATATATATATTTTTATAGCTTA
AAAGTGTGTTATGAATATTTTAAGCATATTTTCTTTTTCTTTGGATTGTGTAAAATGAAC
TCATATAATGCGTTTTTTTGTTTTTGTTATTTTGTCATTTTGTTATTTTGCTATTTTATG
GATTAATTTTTGTTTATAAAATGGGAAATAATTTTAACATATTTAAATAAATGGAGAAAA
AATATAAAATAATTATAAAAAAAAGTTAATACACATTTTTTCCTGTTATAGACCTTATAT
TTATTTATCCATATATATATATATATATATATATATATATATATACATACCAAGTGAATT
AAGAGGAAAGCTAATTTATTATTCAGAATAATATATGAACTATATATAATTTTTATTATT
TTGGTGTATATTAATCTGTCTATATGCATACATGCAATAATTTATCGACTTATATATCAA
ATAACATAAAATAGAAGTGTTTTAAATTATGGATATATGCTCAATATTCATTTTTTTTAA
TAAGTTAGCTATATTTAAATTATACATTTTATATATGGTCTCTTTTTTTTTTAAATATTA
TTTAAGTGATCATGAAAATATAAATAATTTTTTTTTATTTAATATCCTTTTGCTTGCATG
TGGTAAATGGAAATTTGGATGTGTTTTGAARGTTCGGATATAGTTGTATGGACATATAAT
ATATTTTGTGAAAAATTGGTTTTATGTTTATACTTATGCCAATACTTTTTGAGTAAAACA
AAGCAAGTGCTTATAAATAATTAAAGCCAATTTTATAATATATATTTTTTTATTTAATTT
GAATTTAGTAGTATAATTTTTTATGGTAAGTGCTCAAAGAGAGTTGCTTATAAAGTATGG
TTTGTTTCTTTTTCGCCATTTTGAATTACACATTAAAAATATATAGATACATATATTATA
ATATGAAATCATTAATAATTTAGGGAAATTCTACAAATTTAAAAACGAATAAAATAATTG
TTTTTCATCATGCCATAACACAATATTGATATATACATGTACAAACATTTTTTTTATTTG
GAAAATATAAATTATATAAAAAAAAATGTATAGTATACAAAATGAGCATATTCACACGGG
GTGGACGTTCATTTTTTCATTTTTCCCCTGTTTTTTATGAGTATATGATAAAATTTTATG
AACATTTACACAAAATGAAAATGGATATATAGGAAAAATGGAGCGGTATTTCATTTATCT
TTGATTGTCATTTGGATATTATATTACCYTGGGTAGGCAATTAAAAATGTTAAATAACAA
TTTAAGGAAATTATATTTTATATATTAAAATTAACACTGTATTATATGATTCGCTTATAA
AAGCCACTCTTTCCCCATGCAAAGCTGTTTAATATCAATTTTAACAAATTACACACATGT
TAATATATTTATATATATAATTTATATATTTATAATTTATATATTTATATTTTTATTATT
TATATATTTATTATTTATTGTGTGTGTCAATTCGGGTAGGATATACCTCTTTTTTATTGT
TTAAAGCGATTTGTATTCTAAAATATAAAGRATTTGAAAAAGAGAAAGATAGAATATGAT
CCCATCATATATAGCCCTATAATTTTTATTTAGCAGCGAATTAATTTTTCTATTAAGTTT
ATGTGTAATTAAAATAACGGAATATATATAATACAATAAAAAAGTGCATAAATTAAAATT
TTTTCAATTAAATTTTTTTTTTTAAGGGGTTATATAATATTAAATATATAAAATACGATT
ATATATTTTTGCTACAATTTTTTATATTAAGATATAAATAGTAAATAAATGGTATTATAT
GGCATGTAATATATAAATTTTTTCCAATTTTTATTTTATATACACTTTTCCTTTTTTTGT
CATAAAACTTAAACAATTTACACATTCATTTTAAAAATTGACTATTTGTTTCAACATTTT
TTGAGTTTCCGTTTTATAATAGTATTTTCATTTGTATATTGCTTATATATATAAATACAC
ACCTAAATGTTACAAAGGATCAATGCATAAACCGGTGTGTCTGGTCGTCGCGATGACCCC
CAAGAGGGGCATCGGCATCAACAACGGCCTCCCGTGGCCCCACTTGACCACAGATTTCAA
ACACTTTTCTCGTGTGACAAAAACGACGCCCGAAGAAGCCAGTCGCCTGAACGGGTGGCT
TCCCAGGAAATTTGCAAAGACGGGCGACTCTGGACTTCCCTCTCCATCAGTCGGCAAGAG
ATTCAACGCCGTTGTCATGGGACGGAAAACCTGGGAAAGCATGCCTCGAAAGTTTAGACC
CCTCGTGGACAGATTGAACATCGTCGTTTCCTCTTCCCTCAAAGAAGAAGACATTGCGGC
GGAGAAGCCTCAAGCTGAAGGCCAGCAGCGCGTCCGAGTCTGTGCTTCACTCCCAGCAGC
TCTCAGCCTTCTGGAGGAAGAGTACAAGGATTCTGTCGACCAGATTTTTGTCGTGGGAGG
AGCGGGACTGTACGAGGCAGCGCTGTCTCTGGGCGTTGCCTCTCACCTGTACATCACGCG
TGTAGCCCGCGAGTTTCCGTGCGACGTTTTCTTCCCTGCGTTCCCCGGAGATGACATTCT
TTCAAACAAATCAACTGCTGCGCAGGCTGCAGCTCCTGCCGAGTCTGTGTTCGTTCCCTT
TTGTCCGGAGCTCGGAAGAGAGAAGGACAATGAAGCGACGTATCGACCCATCTTCATTTC
CAAGACCTTCTCAGACAACGGGGTTCCCTACGACTTTGTGGTTCTCGAGAAGAGAAGGAA
GACTGACGACGCAGCCACTGCGGAACCGAGCAACGCAATGAGCTCCTTGACGTCCACGAG
GGAGACAACTCCCGTGCACGGGTTGCAGGCTCCTTCTTCGGCCGCAGCCATTGCCCCGGT
GTTGGCGTGGATGGACGAAGAAGACCGGAAAAAACGCGAGCAAAAGGAACTGATTCGGGC
CGTTCCGCATGTTCACTTTAGAGGCCATGAAGAGTTCCAGTACCTTGATCTCATTGCCGA
CATTATTAACAATGGAAGGACAATGGATGACCGAACGGGCGTTGGTGTCATCTCCAAATT
CGGCTGCACTATGCGCTACTCGCTGGATCAGGCCTTTCCACTTCTCACCACAAAGCGTGT
GTTCTGGAAAGGGGTCCTCGAAGAGTTGCTGTGGTTCATTCGCGGCGACACGAACGCAAA
CCATCTTTCTGAGAAGGGCGTGAAGATCTGGGACAAGAATGTGACACGCGAGTTCCTCGA
TTCGCGCAATCTCCCCCACCGAGAGGTCGGAGACATCGGCCCGGGCTACGGCTTCCAGTG
GAGACACTTCGGCGCGGCATACAAAGACATGCACACAGACTACACAGGGCAGGGCGTCGA
CCAGCTGAAGAATGTGATCCAGATGCTGAGAACGAATCCAACAGATCGTCGCATGCTCAT
GACTGCCTGGAATCCTGCAGCGCTGGACGAAATGGCGCTGCCGCCTTGTCACTTGTTGTG
CCAGTTCTACGTGAACGACCAGAAGGAGCTGTCGTGCATCATGTATCAGCGGTCGTGCGA
TGTCGGCCTCGGCGTCCCCTTCAACATCGCTTCCTATTCGCTTTTGACGCTCATGGTTGC
ACACGTCTGCAACCTAAAACCTAAGGAGTTCATTCACTTCATGGGGAACACGCATGTCTA
CACGAACCATGTCGAGGCTTTAAAAGAGCAGCTGCGGAGAGAACCGAGACCGTTCCCCAT
TGTGAACATCCTCAACAAGGAACGCATCAAGGAAATCGACGATTTCACCGCCGAGGATTT
TGAGGTCGTGGGCTACGTCCCGCACGGACGAATCCAGATGGAGATGGCTGTCTAGCGGAA
ATACAGAAGCTAGCTTTGATCCCGTTTTTCTTACTTATATATTTATACCAATTGATTGTA
TTTATAACTGTAAAAATGTGTATGTTGTGTGCATATTTTTTTTTGTGCATGCACATGCAT
GTAAATAGCTAAAATTATGAACATTTTATTTTTTGTTCAGAAAAAAAAAACTTTACACAC
ATAAAATGGCTAGTATGAATAGCCATATTTTATATAAATTAAATCCTATGAATTTATGAC
CATATTAAAAATTTAGATATTTATGGAACATAATATGTTTGAAACAATAAGACAAAATTA
TTATTATTATTATTATTTTTACTGTTATAATTATGTTGTCTCTTCAATGATTCATAAATA
GTTGGACTTGATTTTTAAAATGTTTATAATATGATTAGCATAGTTAAATAAAAAAAGTTG
AAAAATTAAAAAAAAACATATAAACACAAATGATGTTTTTTCCTTCAATTTCGAT
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 PB000198.00.0 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 754 bp fragment (SmaI/ BamHI) of the promoter region of PB000198.00.0.

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_0619200
Gene Model P. falciparum ortholog PF3D7_0721700
Gene productsecreted ookinete protein, putative
Gene product: Alternative namePSOP1, Putative secreted ookinete protein 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 1aaCCCGGGcaatgtgataaaattaagattg
Additional information primer 1forward
Sequence Primer 2aaGGATCCttttatatctgtcttattaagattc
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