Summary

RMgm-126
Malaria parasiteP. berghei
Genotype
Transgene
Transgene not Plasmodium: GFP (gfp-mut3)
Promoter: Gene model: PBANKA_1035200; Gene model (P.falciparum): PF3D7_1407000; Gene product: LCCL domain-containing protein | scavenger receptor-like protein (PbSR, P. berghei Scavenger Receptor-like protein; PSLAP; LAP1, LCCL/lectin adhesive-like 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; Oocyst; Sporozoite;
Last modified: 18 February 2009, 13:46
  *RMgm-126
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
Reference 2 (PMID number) : 18848846
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-126
Principal name516
Alternative namePbCCp3-GFP
Standardized name
Is the mutant parasite cloned after genetic modificationNo
Phenotype
Asexual blood stageNot different from wild type
Gametocyte/GameteGFP expression in female gametocytes. No expression could be detected in asexual blood stages and male gametocytes.
Fertilization and ookineteNot tested
OocystNo GFP expression could be detected in oocysts at day 11 and 17
SporozoiteNo GFP expression could be detected in salivary gland sporozoites
Liver stageNot different from wild type
Additional remarks phenotype

Mutant/mutation
The mutant expresses GFP under the control of the promoter of the gene encoding CCp3 (PbSR, P. berghei Scavenger Receptor-like protein; PSLAP; LAP1;PF14_0067)

Protein (function)
The ccp3 (pbsr, lap1) gene is a member of a small conserved gene family, encoding proteins with multiple adhesive domains, for example a Lgl1 (LCCL)-lectin adhesive domain. In P. falciparum the LCCL domain-containing proteins are termed PfCCp's.
See also the phenotype description of mutants lacking expression of CCp3/PbSR/LAP1 (RMgm-113, RMgm-114) or expressing a GFP- and mCherry-tagged version of PbSR (RMgm-116).

Phenotype
Phenotype analyses indicate absence of promoter activity in asexual blood stages and male gametocytes and active transcription in female gametocytes. Promoter activity appears to be absent in oocysts and sporozoites.  The absence in male gametocytes and prescence in 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 ccp3 (pbsr, lap1) 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 1487 bp fragment (EcoRV/BamHI) of the promoter region of ccp3 (pbsr, lap1).
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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ATCTTTTAAAATAGAAATTCTTTTGGATATGCAATTGACGCAGCATGTGGTCAACTATTT
GCAGGTAATATAAACATCTTTATATATTTCATTTCATACACATTATATGATTCTTACTTA
TATAAAATAATTCTTATCATTGCAATTTTTTATAATTTCCCTATTTATATATCACATTAA
AAATACCATCAATTTTAGACTATGAACCTAAAAAGACAAAGAAAATTACAGGTCCAAAAA
CGTCATTAATAGAATAGTAAAGGTTTGAAAAATTAAATTTGGACTTTATCTTTTTATATG
CAATTATGTAGGAAAATTTATACACATATATATATGTTGAATAAAAAATCAATGTTTTTT
ATTTTAATTTAATCTATATATACATACCCTTTTTTTTTTATTTTTTTTGCACACAATATA
TAGCTAGCTTACCATGCTCATATATTTACTTTTTTGGCTAATCAATTTTTGTTTAATTTT
CCGAAGTTTTAATCAATTTTTAACTTTTTTTATTAACTTTTTTTTGTTATAATTAAAGAA
AATTTTAAAGAGTTTTTTATAAAAAAAAACATTATTCTAAAAATTTGATGTATCTACACG
GCGTTGTTTTTGTTTCGTCCTTCAAATACAACACTTTAAGTTGGTATGCTATGTGCACAT
ACATATATATATAATATTAAATTCGTGCCTCCGTACATTTTTTGTGCCTATTTTAATTAT
TTTTATTGTAATAATTTTAAAAAACAATAACTCACTTTCTTGTATTGCTATATTAAAAAA
TATACATATATTCATTGTATACTTTGATGAAAAATATGTATTATACTTTCTCAATGAGAC
CTAAATAAATGTGTACATATGTTACACTCTATATAAGTATTTTTCTTCATCTTCGTCTTT
TTGTTAAAATAGTCACGTATTTATTCTATACACTATCCATGCACCATTCTATATGTTATA
ATTTACTTATGTCAATTAGCTATTTTTATGAACATCATAAAAAAAATATGTTTATTCTAA
CCAATACATGCAAGAATTCATATATCATTTAAAATGGAATGTGCATAAATGTACAGAAAC
ATATGAAATTTTGTTATTGGGTATTTATTAATAAGAAAAGCATTTGCTTAATTTGGTGAA
TTTTTTGAACGGATTCGCGATGAATAACAACTATCATTATTTGGGTTATTACTTCTTGCA
CACATATATACTGTATATATATATATTAATCAGTTTATTTTTGTATTATAGTTTTTTATA
TATATAGTAAGTTTTTTTTCAGACTAGCATAGTATGCCTTTTTAATCTACAAATTTTTGT
CTAAAAGGTATTTTTAGTGTTCAATATTTTTAACCACATACTAGTGCAAGTTTTTTTTTA
GCTAGCTATTTGTGTGCAATAAATTATTAACAGGGAATAAAAAAAAATAGTTATATAACA
AAATCAAGATACACACTTCCAAACATTTTTTTATATTTTTAGAGAAGAAAGGATCCATGA
GTAAAGGAGAAGAACTTTTCACTGGAGTTGTCCCAATTCTTGTTGAATTAGATGGTGATG
TTAATGGGCACAAATTTTCTGTCAGTGGAGAGGGTGAAGGTGATGCAACATACGGAAAAC
TTACCCTTAAATTTATTTGCACTACTGGAAAACTACCTGTTCCATGGCCAACACTTGTCA
CTACTTTCGGTTATGGTGTTCAATGCTTTGCGAGATACCCAGATCATATGAAACAGCATG
ACTTTTTCAAGAGTGCCATGCCCGAAGGTTATGTACAGGAAAGAACTATATTTTTCAAAG
ATGACGGGAACTACAAGACACGTGCTGAAGTCAAGTTTGAAGGTGATACCCTTGTTAATA
GAATCGAGTTAAAAGGTATTGATTTTAAAGAAGATGGAAACATTCTTGGACACAAATTGG
AATACAACTATAACTCACACAATGTATACATCATGGCAGACAAACAAAAGAATGGAATCA
AAGTTAACTTCAAAATTAGACACAACATTGAAGATGGAAGCGTTCAACTAGCAGACCATT
ATCAACAAAATACTCCAATTGGCGATGGCCCTGTCCTTTTACCAGACAACCATTACCTGT
CCACACAATCTGCCCTTTCGAAAGATCCCAACGAAAAGAGAGACCACATGGTCCTTCTTG
AGTTTGTAACAGCTGCTGGGATTACACATGGCATGGATGAACTATACAAATAAATTCTAG
AAGATCCCGTTTTTCTTACTTATATATTTATACCAATTGATTGTATTTATAACTGTAAAA
ATGTGTATGTTGTGTGCATATTTTTTTTTGTGCATGCACATGCATGTAAATAGCTAAAAT
TATGAACATTTTATTTTTTGTTCAGAAAAAAAAAACTTTACACACATAAAATGGCTAGTA
TGAATAGCCATATTTTATATAAATTAAATCCTATGAATTTATGACCATATTAAAAATTTA
GATATTTATGGAACATAATATGTTTGAAACAATAAGACAAAATTATTATTATTATTATTA
TTTTTACTGTTATAATTATGTTGTCTCTTCAATGATTCATAAATAGTTGGACTTGATTTT
TAAAATGTTTATAATATGATTAGCATAGTTAAATAAAAAAAGTTGAAAAATTAAAAAAAA
ACATATAAACACAAATGATGTTTTTTCCTTCAATTTCGGGTACCGAGCTCGAATTCAACC
TGGTTGATCTTGCCAGTAGTCATATGCTTGTCTCAAAGATTAAGCCATGCAAGTGAAAGT
ATATGCACATTTATTGCAGAAACTGCGAACGGCTCATTAAAACAGTTATAATCTACTTGA
CATTTTATTATAAGGATAACTACGGAAAAGCTGTAGCTAATACTTGTTAAGTACTTTTAC
TCCCCGGAGTAATTGTATGTATTTGTTAAGACCCCTAAGAAAAAATGATATTAAAGGAAT
TATAACAAAGAAGCAACACATAATATAATTATTCAGTGTGTATCAATCGAGTTTCTGACC
TATCAGCTTTTGATGTTAGGGTATTGACCTAACATGGCTTTGACGGGTAACGGGGAATTA
GAGTTCGATTCCGGAGAGGGAGCCTGAGAAATAGCTACCACATCTAAGGAAGGCAGCAGG
CGCGTAAATTACCCAATTCTAAATAAGAGAGGTAGTGACAAGAAATAACAATATAAGGCC
AAATTTTGGTTTTATAATTGGAATGATGGGAATTTAAAACCTTCCCAAAAATCAATTGGA
GGGCAAGTCTGGTGCCAGCAGCCGCGGTAATTCCAGCTCCAATAGCGTATATTAAAATTG
TTGCAGTTAAAACGCTCGTAGTTGAACTTCAAGGGTATAATTATTTTAAGCAACTCACTT
GGAAAGAATCATGACTTCTGTCACTGCTTTTATCCTTGTTGCAGTTCTTTTAATACAGGG
CCCTTTGAGAGCCCATTAATTTATGACTGGGTTTCTCGTTACTTTGAGTAAATTAGAGTG
TTTAAAGCAAACAGATAAAGCGTATTTTACTGTGTTTGAATACTATAGCATGGAATAACA
ACATTGAATAGGTCAAAAGTTTTTGAAAAATTTTTCTTATTTTGGCTTAGATACAGTTAA
TAGGAGTAGCTTGGGGGCATTTGTATTCAGATGTCAGAGGTGAAATTCTTAGATTTTCTG
GAGACAAACAACTGCGAAAGCATTTGCCTAAAATACTTCCATTAATCAAGAACGAAAGTT
AAGGGAGTGAAGACGATCAGATACCGTCGTAATCTTAACCATAAACTATGCCGACTAAGT
GTTGGATGAAAATTTATAAATAAAACTATCTTCTTTAAAGGAGTAGTTTTTTAGATGCTT
CCTTCAGTACCTTATGAGAAATCAAAGTCTTTGGGTTCTGGGGCGAGTATTCGCGCAAGC
GAGAAAGTTAAAAGAATTGACGGAAGGGCACCACCAGGCGTGGAGCTTGCGGCTTAATTT
GACTCAACACGGGGAAACTCACTAGTTTAAGACAAGAGTAGGATTGACAGATTAATAGCT
CTTTCTTGATTTCTTGGATGGTGATGCATGGCCGTTTTTAGTTCGTGAATATGATTTGTC
TGGTTAATTCCGATAACGAACGAGATCTTAACCTGCTAATTAGCGGTGGATATGTGATAT
TCTTCGAAGGTGGACTAACTATAGCGTTTTCGAAGGTATGTTGCATAATCAAATTGGTTT
ACCCTTTGTTTTTTTGTAGCATATTCTTTTATTTCGTTGGGTTTTTTCCCTAGTAAGGAT
GTATCTGCTTTATTTAATGCTTCTTAGAGGAACGATGTGTGTCTAACACAAGGAAGTTTA
AGGCAACAACAGGTCTGTGATGTCCTTAGATATACTAGGCTGCACGCGTGCTACACTGAT
ATGTAAAACGAGTATTTAAAATTATATCTGTATGGTAGATAATTTAATTTCTACGTATTA
TCAGCATATACTTTTCCTACACTGAAATAGTGAAGGTAATCTTTATCAATACATATCGTG
ATGGGGATAGATTATTGCAATTATTAATCTTGAACGAGGAATGCCTAGTAAGCATGATTC
ATCAGATTGTGCTGACTACGTCCCTGCCCTTTGTACACACCGCCCGTCGCTCCTACCGAT
TGAAAGATATGATGAATTGTTTGGACAAGAAAATAGAAATTTTATTTTTATTTTTTTTGG
AAGGACCGTAAATCCTATCTTTTAAAGGAAGGAGAAGTCGTAACAAGGTTTCCGTAGGTG
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
GCATGCCTGCAGGTCAACAATAAATAATAAATAAATATTGTGGAAATAAAATAACATATA
ATTATTTTTAATACATTGATTTCCCTTTTATTTTTTTAAATTTCATTGATATAAAAATAT
ATAATAATAACATATATGATTTCAAATTAATCTTTTCAAAAATGGTGTTTATTTTTGTAT
GTTTGTGTATGAATTAATCACATAACACATCTATTAAATTGAGTTGGTAATATAGACACA
AATAAATATATATATTTTTATAGCTTAAAAGTGTGTTATGAATATTTTAAGCATATTTTC
TTTTTCTTTGGATTGTGTAAAATGAACTCATATAATGCGTTTTTTTGTTTTTGTTATTTT
GTCATTTTGTTATTTTGCTATTTTATGGATTAATTTTTGTTTATAAAATGGGAAATAATT
TTAACATATTTAAATAAATGGAGAAAAAATATAAAATAATTATAAAAAAAAGTTAATACA
CATTTTTTCCTGTTATAGACCTTATATTTATTTATCCATATATATATATATATATATATA
TATATATATATACATACCAAGTGAATTAAGAGGAAAGCTAATTTATTATTCAGAATAATA
TATGAACTATATATAATTTTTATTATTTTGGTGTATATTAATCTGTCTATATGCATACAT
GCAATAATTTATCGACTTATATATCAAATAACATAAAATAGAAGTGTTTTAAATTATGGA
TATATGCTCAATATTCATTTTTTTTAATAAGTTAGCTATATTTAAATTATACATTTTATA
TATGGTCTCTTTTTTTTTTAAATATTATTTAAGTGATCATGAAAATATAAATAATTTTTT
TTTATTTAATATCCTTTTGCTTGCATGTGGTAAATGGAAATTTGGATGTGTTTTGAARGT
TCGGATATAGTTGTATGGACATATAATATATTTTGTGAAAAATTGGTTTTATGTTTATAC
TTATGCCAATACTTTTTGAGTAAAACAAAGCAAGTGCTTATAAATAATTAAAGCCAATTT
TATAATATATATTTTTTTATTTAATTTGAATTTAGTAGTATAATTTTTTATGGTAAGTGC
TCAAAGAGAGTTGCTTATAAAGTATGGTTTGTTTCTTTTTCGCCATTTTGAATTACACAT
TAAAAATATATAGATACATATATTATAATATGAAATCATTAATAATTTAGGGAAATTCTA
CAAATTTAAAAACGAATAAAATAATTGTTTTTCATCATGCCATAACACAATATTGATATA
TACATGTACAAACATTTTTTTTATTTGGAAAATATAAATTATATAAAAAAAAATGTATAG
TATACAAAATGAGCATATTCACACGGGGTGGACGTTCATTTTTTCATTTTTCCCCTGTTT
TTTATGAGTATATGATAAAATTTTATGAACATTTACACAAAATGAAAATGGATATATAGG
AAAAATGGAGCGGTATTTCATTTATCTTTGATTGTCATTTGGATATTATATTACCYTGGG
TAGGCAATTAAAAATGTTAAATAACAATTTAAGGAAATTATATTTTATATATTAAAATTA
ACACTGTATTATATGATTCGCTTATAAAAGCCACTCTTTCCCCATGCAAAGCTGTTTAAT
ATCAATTTTAACAAATTACACACATGTTAATATATTTATATATATAATTTATATATTTAT
AATTTATATATTTATATTTTTATTATTTATATATTTATTATTTATTGTGTGTGTCAATTC
GGGTAGGATATACCTCTTTTTTATTGTTTAAAGCGATTTGTATTCTAAAATATAAAGRAT
TTGAAAAAGAGAAAGATAGAATATGATCCCATCATATATAGCCCTATAATTTTTATTTAG
CAGCGAATTAATTTTTCTATTAAGTTTATGTGTAATTAAAATAACGGAATATATATAATA
CAATAAAAAAGTGCATAAATTAAAATTTTTTCAATTAAATTTTTTTTTTTAAGGGGTTAT
ATAATATTAAATATATAAAATACGATTATATATTTTTGCTACAATTTTTTATATTAAGAT
ATAAATAGTAAATAAATGGTATTATATGGCATGTAATATATAAATTTTTTCCAATTTTTA
TTTTATATACACTTTTCCTTTTTTTGTCATAAAACTTAAACAATTTACACATTCATTTTA
AAAATTGACTATTTGTTTCAACATTTTTTGAGTTTCCGTTTTATAATAGTATTTTCATTT
GTATATTGCTTATATATATAAATACACACCTAAATGTTACAAAGGATCAATGCATAAACC
GGTGTGTCTGGTCGTCGCGATGACCCCCAAGAGGGGCATCGGCATCAACAACGGCCTCCC
GTGGCCCCACTTGACCACAGATTTCAAACACTTTTCTCGTGTGACAAAAACGACGCCCGA
AGAAGCCAGTCGCCTGAACGGGTGGCTTCCCAGGAAATTTGCAAAGACGGGCGACTCTGG
ACTTCCCTCTCCATCAGTCGGCAAGAGATTCAACGCCGTTGTCATGGGACGGAAAACCTG
GGAAAGCATGCCTCGAAAGTTTAGACCCCTCGTGGACAGATTGAACATCGTCGTTTCCTC
TTCCCTCAAAGAAGAAGACATTGCGGCGGAGAAGCCTCAAGCTGAAGGCCAGCAGCGCGT
CCGAGTCTGTGCTTCACTCCCAGCAGCTCTCAGCCTTCTGGAGGAAGAGTACAAGGATTC
TGTCGACCAGATTTTTGTCGTGGGAGGAGCGGGACTGTACGAGGCAGCGCTGTCTCTGGG
CGTTGCCTCTCACCTGTACATCACGCGTGTAGCCCGCGAGTTTCCGTGCGACGTTTTCTT
CCCTGCGTTCCCCGGAGATGACATTCTTTCAAACAAATCAACTGCTGCGCAGGCTGCAGC
TCCTGCCGAGTCTGTGTTCGTTCCCTTTTGTCCGGAGCTCGGAAGAGAGAAGGACAATGA
AGCGACGTATCGACCCATCTTCATTTCCAAGACCTTCTCAGACAACGGGGTTCCCTACGA
CTTTGTGGTTCTCGAGAAGAGAAGGAAGACTGACGACGCAGCCACTGCGGAACCGAGCAA
CGCAATGAGCTCCTTGACGTCCACGAGGGAGACAACTCCCGTGCACGGGTTGCAGGCTCC
TTCTTCGGCCGCAGCCATTGCCCCGGTGTTGGCGTGGATGGACGAAGAAGACCGGAAAAA
ACGCGAGCAAAAGGAACTGATTCGGGCCGTTCCGCATGTTCACTTTAGAGGCCATGAAGA
GTTCCAGTACCTTGATCTCATTGCCGACATTATTAACAATGGAAGGACAATGGATGACCG
AACGGGCGTTGGTGTCATCTCCAAATTCGGCTGCACTATGCGCTACTCGCTGGATCAGGC
CTTTCCACTTCTCACCACAAAGCGTGTGTTCTGGAAAGGGGTCCTCGAAGAGTTGCTGTG
GTTCATTCGCGGCGACACGAACGCAAACCATCTTTCTGAGAAGGGCGTGAAGATCTGGGA
CAAGAATGTGACACGCGAGTTCCTCGATTCGCGCAATCTCCCCCACCGAGAGGTCGGAGA
CATCGGCCCGGGCTACGGCTTCCAGTGGAGACACTTCGGCGCGGCATACAAAGACATGCA
CACAGACTACACAGGGCAGGGCGTCGACCAGCTGAAGAATGTGATCCAGATGCTGAGAAC
GAATCCAACAGATCGTCGCATGCTCATGACTGCCTGGAATCCTGCAGCGCTGGACGAAAT
GGCGCTGCCGCCTTGTCACTTGTTGTGCCAGTTCTACGTGAACGACCAGAAGGAGCTGTC
GTGCATCATGTATCAGCGGTCGTGCGATGTCGGCCTCGGCGTCCCCTTCAACATCGCTTC
CTATTCGCTTTTGACGCTCATGGTTGCACACGTCTGCAACCTAAAACCTAAGGAGTTCAT
TCACTTCATGGGGAACACGCATGTCTACACGAACCATGTCGAGGCTTTAAAAGAGCAGCT
GCGGAGAGAACCGAGACCGTTCCCCATTGTGAACATCCTCAACAAGGAACGCATCAAGGA
AATCGACGATTTCACCGCCGAGGATTTTGAGGTCGTGGGCTACGTCCCGCACGGACGAAT
CCAGATGGAGATGGCTGTCTAGCGGAAATACAGAAGCTAGCTTTGATCCCGTTTTTCTTA
CTTATATATTTATACCAATTGATTGTATTTATAACTGTAAAAATGTGTATGTTGTGTGCA
TATTTTTTTTTGTGCATGCACATGCATGTAAATAGCTAAAATTATGAACATTTTATTTTT
TGTTCAGAAAAAAAAAACTTTACACACATAAAATGGCTAGTATGAATAGCCATATTTTAT
ATAAATTAAATCCTATGAATTTATGACCATATTAAAAATTTAGATATTTATGGAACATAA
TATGTTTGAAACAATAAGACAAAATTATTATTATTATTATTATTTTTACTGTTATAATTA
TGTTGTCTCTTCAATGATTCATAAATAGTTGGACTTGATTTTTAAAATGTTTATAATATG
ATTAGCATAGTTAAATAAAAAAAGTTGAAAAATTAAAAAAAAACATATAAACACAAATGA
TGTTTTTTCCTTCAATTTCGAT
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 ccp3 (pbsr, lap1) 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 1487 bp fragment (EcoRV/BamHI) of the promoter region of ccp3 (pbsr, lap1).

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_1035200
Gene Model P. falciparum ortholog PF3D7_1407000
Gene productLCCL domain-containing protein | scavenger receptor-like protein
Gene product: Alternative namePbSR, P. berghei Scavenger Receptor-like protein; PSLAP; LAP1, LCCL/lectin adhesive-like 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 1aaGATATCttttaaaatagaaattcttttgg
Additional information primer 1forward
Sequence Primer 2aaGGATCCtttcttctctaaaaatataaaaaaatgtttgg
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