RMgmDB - Rodent Malaria genetically modified Parasites

Summary

RMgm-131

Malaria parasite	P. 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 modification	The 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
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Parent parasite used to introduce the genetic modification
Rodent Malaria Parasite	P. berghei
Parent strain/line	P. berghei ANKA
Name parent line/clone	P. berghei ANKA cl15cy1
Other information parent line	A reference wild type clone from the ANKA strain of P. berghei (PubMed: PMID: 17406255).
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The mutant parasite was generated by
Name PI/Researcher	S.M. Khan, G.R. Mair, C.J. Janse, A.P. Waters
Name Group/Department	Leiden Malaria Research Group
Name Institute	Leiden University Medical Center (LUMC)
City	Leiden
Country	The Netherlands
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Name of the mutant parasite
RMgm number	RMgm-131
Principal name	500
Alternative name
Standardized name
Is the mutant parasite cloned after genetic modification	No
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Phenotype
Asexual blood stage	Not different from wild type
Gametocyte/Gamete	GFP expression in male gametocytes. No expression could be detected in asexual blood stages and female gametocytes.
Fertilization and ookinete	Not tested
Oocyst	Not tested
Sporozoite	Not tested
Liver stage	Not 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

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Type and details of transgene

Is the transgene Plasmodium derived Transgene: not Plasmodium

Transgene name GFP (gfp-mut3)

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Details of the genetic modification

Inducable system used No

Additional remarks inducable system

Type of plasmid/construct Plasmid single cross-over

PlasmoGEM (Sanger) construct/vector used No

Modified PlasmoGEM construct/vector used No

Plasmid/construct map

Plasmid/construct sequence

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 parasite tgdhfr

Promoter of the selectable marker pbdhfr

Selection (positive) procedure pyrimethamine

Selection (negative) procedure No

Additional remarks genetic modification 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.

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

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Other details transgene

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Promoter

Gene Model of Parasite PBANKA_0925400

Gene Model P. falciparum ortholog PF3D7_1122900

Gene product dynein 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 1	aaGTTAACacacgtaaatgttttaatgcac
Additional information primer 1	forward
Sequence Primer 2	aaGGATCCattttatacaaacctatatatacac
Additional information primer 2	reverse

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3'-UTR

Gene Model of Parasite PBANKA_0719300

Gene product bifunctional dihydrofolate reductase-thymidylate synthase, putative

Gene product: Alternative name dhfr/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 / Insertion Insertion locus

Gene Model of Parasite Not available

Gene product Not available

Gene product: Alternative name small 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

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