Summary

RMgm-128
Malaria parasiteP. berghei
Genotype
Transgene
Transgene not Plasmodium: GFP (gfp-mut3)
Promoter: Gene model: PBANKA_0417600; Gene model (P.falciparum): PF3D7_0903800; Gene product: LCCL domain-containing protein (LAP6, LCCL/lectin adhesive-like protein 2; CCp4)
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, 13:49
  *RMgm-128
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) : 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/ResearcherC. Lavazec, G.R. Mair, C.J. Janse, J. Templeton
Name Group/DepartmentDepartment of Microbiology and Immunology
Name InstituteWeill Cornell Medical College
CityNew York
CountryUSA
Name of the mutant parasite
RMgm numberRMgm-128
Principal name1014
Alternative namePbCCp4-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 LAP6 (LCCL/lectin adhesive-like protein 2; LCCL domain containing protein CCp4; PFI0185w).

Protein (function)
The lap6/ccp4 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 a mutants lacking expression of LAP6/CCp4 (RMgm-120, RMgm-125).

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 (see also '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 lap6/ccp4  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 650 bp fragment of the promoter region of lap6/ccp4.
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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TATGCTTGTCTCAAAGATTAAGCCATGCAAGTGAAAGTATATGCACATTTATTGCAGAAA
CTGCGAACGGCTCATTAAAACAGTTATAATCTACTTGACATTTTATTATAAGGATAACTA
CGGAAAAGCTGTAGCTAATACTTGTTAAGTACTTTTACTCCCCGGAGTAATTGTATGTAT
TTGTTAAGACCCCTAAGAAAAAATGATATTAAAGGAATTATAACAAAGAAGCAACACATA
ATATAATTATTCAGTGTGTATCAATCGAGTTTCTGACCTATCAGCTTTTGATGTTAGGGT
ATTGACCTAACATGGCTTTGACGGGTAACGGGGAATTAGAGTTCGATTCCGGAGAGGGAG
CCTGAGAAATAGCTACCACATCTAAGGAAGGCAGCAGGCGCGTAAATTACCCAATTCTAA
ATAAGAGAGGTAGTGACAAGAAATAACAATATAAGGCCAAATTTTGGTTTTATAATTGGA
ATGATGGGAATTTAAAACCTTCCCAAAAATCAATTGGAGGGCAAGTCTGGTGCCAGCAGC
CGCGGTAATTCCAGCTCCAATAGCGTATATTAAAATTGTTGCAGTTAAAACGCTCGTAGT
TGAACTTCAAGGGTATAATTATTTTAAGCAACTCACTTGGAAAGAATCATGACTTCTGTC
ACTGCTTTTATCCTTGTTGCAGTTCTTTTAATACAGGGCCCTTTGAGAGCCCATTAATTT
ATGACTGGGTTTCTCGTTACTTTGAGTAAATTAGAGTGTTTAAAGCAAACAGATAAAGCG
TATTTTACTGTGTTTGAATACTATAGCATGGAATAACAACATTGAATAGGTCAAAAGTTT
TTGAAAAATTTTTCTTATTTTGGCTTAGATACAGTTAATAGGAGTAGCTTGGGGGCATTT
GTATTCAGATGTCAGAGGTGAAATTCTTAGATTTTCTGGAGACAAACAACTGCGAAAGCA
TTTGCCTAAAATACTTCCATTAATCAAGAACGAAAGTTAAGGGAGTGAAGACGATCAGAT
ACCGTCGTAATCTTAACCATAAACTATGCCGACTAAGTGTTGGATGAAAATTTATAAATA
AAACTATCTTCTTTAAAGGAGTAGTTTTTTAGATGCTTCCTTCAGTACCTTATGAGAAAT
CAAAGTCTTTGGGTTCTGGGGCGAGTATTCGCGCAAGCGAGAAAGTTAAAAGAATTGACG
GAAGGGCACCACCAGGCGTGGAGCTTGCGGCTTAATTTGACTCAACACGGGGAAACTCAC
TAGTTTAAGACAAGAGTAGGATTGACAGATTAATAGCTCTTTCTTGATTTCTTGGATGGT
GATGCATGGCCGTTTTTAGTTCGTGAATATGATTTGTCTGGTTAATTCCGATAACGAACG
AGATCTTAACCTGCTAATTAGCGGTGGATATGTGATATTCTTCGAAGGTGGACTAACTAT
AGCGTTTTCGAAGGTATGTTGCATAATCAAATTGGTTTACCCTTTGTTTTTTTGTAGCAT
ATTCTTTTATTTCGTTGGGTTTTTTCCCTAGTAAGGATGTATCTGCTTTATTTAATGCTT
CTTAGAGGAACGATGTGTGTCTAACACAAGGAAGTTTAAGGCAACAACAGGTCTGTGATG
TCCTTAGATATACTAGGCTGCACGCGTGCTACACTGATATGTAAAACGAGTATTTAAAAT
TATATCTGTATGGTAGATAATTTAATTTCTACGTATTATCAGCATATACTTTTCCTACAC
TGAAATAGTGAAGGTAATCTTTATCAATACATATCGTGATGGGGATAGATTATTGCAATT
ATTAATCTTGAACGAGGAATGCCTAGTAAGCATGATTCATCAGATTGTGCTGACTACGTC
CCTGCCCTTTGTACACACCGCCCGTCGCTCCTACCGATTGAAAGATATGATGAATTGTTT
GGACAAGAAAATAGAAATTTTATTTTTATTTTTTTTGGAAGGACCGTAAATCCTATCTTT
TAAAGGAAGGAGAAGTCGTAACAAGGTTTCCGTAGGTGAATTCACTGGCCGTCGTTTTAC
AACGTCGTGACTGGGAAAACCCTGGCGTTACCCAACTTAATCGCCTTGCAGCACATCCCC
CTTTCGCCAGCTGGCGTAATAGCGAAGAGGCCCGCACCGATCGCCCTTCCCAACAGTTGC
GCAGCCTGAATGGCGAATGGCGCCTGATGCGGTATTTTCTCCTTACGCATCTGTGCGGTA
TTTCACACCGCATATGGTGCACTCTCAGTACAATCTGCTCTGATGCCGCATAGTTAAGCC
AGCCCCGACACCCGCCAACACCCGCTGACGCGCCCTGACGGGCTTGTCTGCTCCCGGCAT
CCGCTTACAGACAAGCTGTGACCGTCTCCGGGAGCTGCATGTGTCAGAGGTTTTCACCGT
CATCACCGAAACGCGCGAGACGAAAGGGCCTCGTGATACGCCTATTTTTATAGGTTAATG
TCATGATAATAATGGTTTCTTAGACGTCAGGTGGCACTTTTCGGGGAAATGTGCGCGGAA
CCCCTATTTGTTTATTTTTCTAAATACATTCAAATATGTATCCGCTCATGAGACAATAAC
CCTGATAAATGCTTCAATAATATTGAAAAAGGAAGAGTATGAGTATTCAACATTTCCGTG
TCGCCCTTATTCCCTTTTTTGCGGCATTTTGCCTTCCTGTTTTTGCTCACCCAGAAACGC
TGGTGAAAGTAAAAGATGCTGAAGATCAGTTGGGTGCACGAGTGGGTTACATCGAACTGG
ATCTCAACAGCGGTAAGATCCTTGAGAGTTTTCGCCCCGAAGAACGTTTTCCAATGATGA
GCACTTTTAAAGTTCTGCTATGTGGCGCGGTATTATCCCGTATTGACGCCGGGCAAGAGC
AACTCGGTCGCCGCATACACTATTCTCAGAATGACTTGGTTGAGTACTCACCAGTCACAG
AAAAGCATCTTACGGATGGCATGACAGTAAGAGAATTATGCAGTGCTGCCATAACCATGA
GTGATAACACTGCGGCCAACTTACTTCTGACAACGATCGGAGGACCGAAGGAGCTAACCG
CTTTTTTGCACAACATGGGGGATCATGTAACTCGCCTTGATCGTTGGGAACCGGAGCTGA
ATGAAGCCATACCAAACGACGAGCGTGACACCACGATGCCTGTAGCAATGGCAACAACGT
TGCGCAAACTATTAACTGGCGAACTACTTACTCTAGCTTCCCGGCAACAATTAATAGACT
GGATGGAGGCGGATAAAGTTGCAGGACCACTTCTGCGCTCGGCCCTTCCGGCTGGCTGGT
TTATTGCTGATAAATCTGGAGCCGGTGAGCGTGGGTCTCGCGGTATCATTGCAGCACTGG
GGCCAGATGGTAAGCCCTCCCGTATCGTAGTTATCTACACGACGGGGAGTCAGGCAACTA
TGGATGAACGAAATAGACAGATCGCTGAGATAGGTGCCTCACTGATTAAGCATTGGTAAC
TGTCAGACCAAGTTTACTCATATATACTTTAGATTGATTTAAAACTTCATTTTTAATTTA
AAAGGATCTAGGTGAAGATCCTTTTTGATAATCTCATGACCAAAATCCCTTAACGTGAGT
TTTCGTTCCACTGAGCGTCAGACCCCGTAGAAAAGATCAAAGGATCTTCTTGAGATCCTT
TTTTTCTGCGCGTAATCTGCTGCTTGCAAACAAAAAAACCACCGCTACCAGCGGTGGTTT
GTTTGCCGGATCAAGAGCTACCAACTCTTTTTCCGAAGGTAACTGGCTTCAGCAGAGCGC
AGATACCAAATACTGTCCTTCTAGTGTAGCCGTAGTTAGGCCACCACTTCAAGAACTCTG
TAGCACCGCCTACATACCTCGCTCTGCTAATCCTGTTACCAGTGGCTGCTGCCAGTGGCG
ATAAGTCGTGTCTTACCGGGTTGGACTCAAGACGATAGTTACCGGATAAGGCGCAGCGGT
CGGGCTGAACGGGGGGTTCGTGCACACAGCCCAGCTTGGAGCGAACGACCTACACCGAAC
TGAGATACCTACAGCGTGAGCATTGAGAAAGCGCCACGCTTCCCGAAGGGAGAAAGGCGG
ACAGGTATCCGGTAAGCGGCAGGGTCGGAACAGGAGAGCGCACGAGGGAGCTTCCAGGGG
GAAACGCCTGGTATCTTTATAGTCCTGTCGGGTTTCGCCACCTCTGACTTGAGCGTCGAT
TTTTGTGATGCTCGTCAGGGGGGCGGAGCCTATGGAAAAACGCCAGCAACGCGGCCTTTT
TACGGTTCCTGGCCTTTTGCTGGCCTTTTGCTCACATGTTCTTTCCTGCGTTATCCCCTG
ATTCTGTGGATAACCGTATTACCGCCTTTGAGTGAGCTGATACCGCTCGCCGCAGCCGAA
CGACCGAGCGCAGCGAGTCAGTGAGCGAGGAAGCGGAAGAGCGCCCAATACGCAAACCGC
CTCTCCCCGCGCGTTGGCCGATTCATTAATGCAGCTGGCACGACAGGTTTCCCGACTGGA
AAGCGGGCAGTGAGCGCAACGCAATTAATGTGAGTTAGCTCACTCATTAGGCACCCCAGG
CTTTACACTTTATGCTTCCGGCTCGTATGTTGTGTGGAATTGTGAGCGGATAACAATTTC
ACACAGGAAACAGCTATGACCATGATTACGCCAAGCTTGCATGCCTGCAGGTCAACAATA
AATAATAAATAAATATTGTGGAAATAAAATAACATATAATTATTTTTAATACATTGATTT
CCCTTTTATTTTTTTAAATTTCATTGATATAAAAATATATAATAATAACATATATGATTT
CAAATTAATCTTTTCAAAAATGGTGTTTATTTTTGTATGTTTGTGTATGAATTAATCACA
TAACACATCTATTAAATTGAGTTGGTAATATAGACACAAATAAATATATATATTTTTATA
GCTTAAAAGTGTGTTATGAATATTTTAAGCATATTTTCTTTTTCTTTGGATTGTGTAAAA
TGAACTCATATAATGCGTTTTTTTGTTTTTGTTATTTTGTCATTTTGTTATTTTGCTATT
TTATGGATTAATTTTTGTTTATAAAATGGGAAATAATTTTAACATATTTAAATAAATGGA
GAAAAAATATAAAATAATTATAAAAAAAAGTTAATACACATTTTTTCCTGTTATAGACCT
TATATTTATTTATCCATATATATATATATATATATATATATATATATATACATACCAAGT
GAATTAAGAGGAAAGCTAATTTATTATTCAGAATAATATATGAACTATATATAATTTTTA
TTATTTTGGTGTATATTAATCTGTCTATATGCATACATGCAATAATTTATCGACTTATAT
ATCAAATAACATAAAATAGAAGTGTTTTAAATTATGGATATATGCTCAATATTCATTTTT
TTTAATAAGTTAGCTATATTTAAATTATACATTTTATATATGGTCTCTTTTTTTTTTAAA
TATTATTTAAGTGATCATGAAAATATAAATAATTTTTTTTTATTTAATATCCTTTTGCTT
GCATGTGGTAAATGGAAATTTGGATGTGTTTTGAARGTTCGGATATAGTTGTATGGACAT
ATAATATATTTTGTGAAAAATTGGTTTTATGTTTATACTTATGCCAATACTTTTTGAGTA
AAACAAAGCAAGTGCTTATAAATAATTAAAGCCAATTTTATAATATATATTTTTTTATTT
AATTTGAATTTAGTAGTATAATTTTTTATGGTAAGTGCTCAAAGAGAGTTGCTTATAAAG
TATGGTTTGTTTCTTTTTCGCCATTTTGAATTACACATTAAAAATATATAGATACATATA
TTATAATATGAAATCATTAATAATTTAGGGAAATTCTACAAATTTAAAAACGAATAAAAT
AATTGTTTTTCATCATGCCATAACACAATATTGATATATACATGTACAAACATTTTTTTT
ATTTGGAAAATATAAATTATATAAAAAAAAATGTATAGTATACAAAATGAGCATATTCAC
ACGGGGTGGACGTTCATTTTTTCATTTTTCCCCTGTTTTTTATGAGTATATGATAAAATT
TTATGAACATTTACACAAAATGAAAATGGATATATAGGAAAAATGGAGCGGTATTTCATT
TATCTTTGATTGTCATTTGGATATTATATTACCYTGGGTAGGCAATTAAAAATGTTAAAT
AACAATTTAAGGAAATTATATTTTATATATTAAAATTAACACTGTATTATATGATTCGCT
TATAAAAGCCACTCTTTCCCCATGCAAAGCTGTTTAATATCAATTTTAACAAATTACACA
CATGTTAATATATTTATATATATAATTTATATATTTATAATTTATATATTTATATTTTTA
TTATTTATATATTTATTATTTATTGTGTGTGTCAATTCGGGTAGGATATACCTCTTTTTT
ATTGTTTAAAGCGATTTGTATTCTAAAATATAAAGRATTTGAAAAAGAGAAAGATAGAAT
ATGATCCCATCATATATAGCCCTATAATTTTTATTTAGCAGCGAATTAATTTTTCTATTA
AGTTTATGTGTAATTAAAATAACGGAATATATATAATACAATAAAAAAGTGCATAAATTA
AAATTTTTTCAATTAAATTTTTTTTTTTAAGGGGTTATATAATATTAAATATATAAAATA
CGATTATATATTTTTGCTACAATTTTTTATATTAAGATATAAATAGTAAATAAATGGTAT
TATATGGCATGTAATATATAAATTTTTTCCAATTTTTATTTTATATACACTTTTCCTTTT
TTTGTCATAAAACTTAAACAATTTACACATTCATTTTAAAAATTGACTATTTGTTTCAAC
ATTTTTTGAGTTTCCGTTTTATAATAGTATTTTCATTTGTATATTGCTTATATATATAAA
TACACACCTAAATGTTACAAAGGATCAATGCATAAACCGGTGTGTCTGGTCGTCGCGATG
ACCCCCAAGAGGGGCATCGGCATCAACAACGGCCTCCCGTGGCCCCACTTGACCACAGAT
TTCAAACACTTTTCTCGTGTGACAAAAACGACGCCCGAAGAAGCCAGTCGCCTGAACGGG
TGGCTTCCCAGGAAATTTGCAAAGACGGGCGACTCTGGACTTCCCTCTCCATCAGTCGGC
AAGAGATTCAACGCCGTTGTCATGGGACGGAAAACCTGGGAAAGCATGCCTCGAAAGTTT
AGACCCCTCGTGGACAGATTGAACATCGTCGTTTCCTCTTCCCTCAAAGAAGAAGACATT
GCGGCGGAGAAGCCTCAAGCTGAAGGCCAGCAGCGCGTCCGAGTCTGTGCTTCACTCCCA
GCAGCTCTCAGCCTTCTGGAGGAAGAGTACAAGGATTCTGTCGACCAGATTTTTGTCGTG
GGAGGAGCGGGACTGTACGAGGCAGCGCTGTCTCTGGGCGTTGCCTCTCACCTGTACATC
ACGCGTGTAGCCCGCGAGTTTCCGTGCGACGTTTTCTTCCCTGCGTTCCCCGGAGATGAC
ATTCTTTCAAACAAATCAACTGCTGCGCAGGCTGCAGCTCCTGCCGAGTCTGTGTTCGTT
CCCTTTTGTCCGGAGCTCGGAAGAGAGAAGGACAATGAAGCGACGTATCGACCCATCTTC
ATTTCCAAGACCTTCTCAGACAACGGGGTTCCCTACGACTTTGTGGTTCTCGAGAAGAGA
AGGAAGACTGACGACGCAGCCACTGCGGAACCGAGCAACGCAATGAGCTCCTTGACGTCC
ACGAGGGAGACAACTCCCGTGCACGGGTTGCAGGCTCCTTCTTCGGCCGCAGCCATTGCC
CCGGTGTTGGCGTGGATGGACGAAGAAGACCGGAAAAAACGCGAGCAAAAGGAACTGATT
CGGGCCGTTCCGCATGTTCACTTTAGAGGCCATGAAGAGTTCCAGTACCTTGATCTCATT
GCCGACATTATTAACAATGGAAGGACAATGGATGACCGAACGGGCGTTGGTGTCATCTCC
AAATTCGGCTGCACTATGCGCTACTCGCTGGATCAGGCCTTTCCACTTCTCACCACAAAG
CGTGTGTTCTGGAAAGGGGTCCTCGAAGAGTTGCTGTGGTTCATTCGCGGCGACACGAAC
GCAAACCATCTTTCTGAGAAGGGCGTGAAGATCTGGGACAAGAATGTGACACGCGAGTTC
CTCGATTCGCGCAATCTCCCCCACCGAGAGGTCGGAGACATCGGCCCGGGCTACGGCTTC
CAGTGGAGACACTTCGGCGCGGCATACAAAGACATGCACACAGACTACACAGGGCAGGGC
GTCGACCAGCTGAAGAATGTGATCCAGATGCTGAGAACGAATCCAACAGATCGTCGCATG
CTCATGACTGCCTGGAATCCTGCAGCGCTGGACGAAATGGCGCTGCCGCCTTGTCACTTG
TTGTGCCAGTTCTACGTGAACGACCAGAAGGAGCTGTCGTGCATCATGTATCAGCGGTCG
TGCGATGTCGGCCTCGGCGTCCCCTTCAACATCGCTTCCTATTCGCTTTTGACGCTCATG
GTTGCACACGTCTGCAACCTAAAACCTAAGGAGTTCATTCACTTCATGGGGAACACGCAT
GTCTACACGAACCATGTCGAGGCTTTAAAAGAGCAGCTGCGGAGAGAACCGAGACCGTTC
CCCATTGTGAACATCCTCAACAAGGAACGCATCAAGGAAATCGACGATTTCACCGCCGAG
GATTTTGAGGTCGTGGGCTACGTCCCGCACGGACGAATCCAGATGGAGATGGCTGTCTAG
CGGAAATACAGAAGCTAGCTTTGATCCCGTTTTTCTTACTTATATATTTATACCAATTGA
TTGTATTTATAACTGTAAAAATGTGTATGTTGTGTGCATATTTTTTTTTGTGCATGCACA
TGCATGTAAATAGCTAAAATTATGAACATTTTATTTTTTGTTCAGAAAAAAAAAACTTTA
CACACATAAAATGGCTAGTATGAATAGCCATATTTTATATAAATTAAATCCTATGAATTT
ATGACCATATTAAAAATTTAGATATTTATGGAACATAATATGTTTGAAACAATAAGACAA
AATTATTATTATTATTATTATTTTTACTGTTATAATTATGTTGTCTCTTCAATGATTCAT
AAATAGTTGGACTTGATTTTTAAAATGTTTATAATATGATTAGCATAGTTAAATAAAAAA
AGTTGAAAAATTAAAAAAAAACATATAAACACAAATGATGTTTTTTCCTTCAATTTCGAT
ATCAGAGAAAATGGCAATTCATAAAATAAAAAAAAGTATATAGCATATTTTATCATACTC
ATGAAAATATTTACGCATACGCTTTGTGAGCATATAATTTATAAAATTGAAAAAAGCATT
GCATATATTTATTTTTATATTGTGACAACAACATATCGGTCCATACCATTTTTTAGACAT
TACTTTATTTTACTATTTTCTATTATTATTACAGTTTTATTTTTCCTCTCCTGCATCTAC
AATAATATTTAATAATATATAAAAATAAAATTGTGTAGATACACATGGGAACATGGATAT
TCTTCTCTATAATAACATGTATTTCGGTAAAAGAATTTAATTTTTTTATGCTTAAAATAT
TATATCCTGTTGTATATAAAATAACTTCTACATATTCTAATATTTAAATAAATTTATAAT
AAAACTTTATCTTATATTTATTTAGTTTGTCTTTGTTTTATCTCTATACTTATATCCACT
TATCTTTTGCTTCTAAACTTTTTGTTCATTCTCATTTTGTTCTGGGACTTTTTTGATTGA
ATAATTTTAATTAATTATGTATTAACATTGTTATTAATATTGTTAATTTAATTATCTTTT
TGATATAATTTTAAAAAGAACAAAGAAAAATATTTACACACAATGGATCCATGAGTAAAG
GAGAAGAACTTTTCACTGGAGTTGTCCCAATTCTTGTTGAATTAGATGGTGATGTTAATG
GGCACAAATTTTCTGTCAGTGGAGAGGGTGAAGGTGATGCAACATACGGAAAACTTACCC
TTAAATTTATTTGCACTACTGGAAAACTACCTGTTCCATGGCCAACACTTGTCACTACTT
TCGGTTATGGTGTTCAATGCTTTGCGAGATACCCAGATCATATGAAACAGCATGACTTTT
TCAAGAGTGCCATGCCCGAAGGTTATGTACAGGAAAGAACTATATTTTTCAAAGATGACG
GGAACTACAAGACACGTGCTGAAGTCAAGTTTGAAGGTGATACCCTTGTTAATAGAATCG
AGTTAAAAGGTATTGATTTTAAAGAAGATGGAAACATTCTTGGACACAAATTGGAATACA
ACTATAACTCACACAATGTATACATCATGGCAGACAAACAAAAGAATGGAATCAAAGTTA
ACTTCAAAATTAGACACAACATTGAAGATGGAAGCGTTCAACTAGCAGACCATTATCAAC
AAAATACTCCAATTGGCGATGGCCCTGTCCTTTTACCAGACAACCATTACCTGTCCACAC
AATCTGCCCTTTCGAAAGATCCCAACGAAAAGAGAGACCACATGGTCCTTCTTGAGTTTG
TAACAGCTGCTGGGATTACACATGGCATGGATGAACTATACAAATAAATTCTAGAAGATC
CCGTTTTTCTTACTTATATATTTATACCAATTGATTGTATTTATAACTGTAAAAATGTGT
ATGTTGTGTGCATATTTTTTTTTGTGCATGCACATGCATGTAAATAGCTAAAATTATGAA
CATTTTATTTTTTGTTCAGAAAAAAAAAACTTTACACACATAAAATGGCTAGTATGAATA
GCCATATTTTATATAAATTAAATCCTATGAATTTATGACCATATTAAAAATTTAGATATT
TATGGAACATAATATGTTTGAAACAATAAGACAAAATTATTATTATTATTATTATTTTTA
CTGTTATAATTATGTTGTCTCTTCAATGATTCATAAATAGTTGGACTTGATTTTTAAAAT
GTTTATAATATGATTAGCATAGTTAAATAAAAAAAGTTGAAAAATTAAAAAAAAACATAT
AAACACAAATGATGTTTTTTCCTTCAATTTCGGGTACCGAGCTCGAATTCAACCTGGTTG
ATCTTGCCAGTAGTCA
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 lap6/ccp4 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 650 bp fragment of the promoter region of lap6/ccp4.

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_0417600
Gene Model P. falciparum ortholog PF3D7_0903800
Gene productLCCL domain-containing protein
Gene product: Alternative nameLAP6, LCCL/lectin adhesive-like protein 2; CCp4
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 15′-AAGATATCAGAGAAAATGGC
Additional information primer 1forward
Sequence Primer 25′-CCGGATCCATTGTGTGTAAATATTTTTCTTTG
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