Summary

RMgm-28
Malaria parasiteP. berghei
Genotype
Transgene
Transgene not Plasmodium: A fusion of GFP (gfp-65) and Luciferase Firefly (LucIAV)
Promoter: Gene model: PBANKA_1133300; Gene model (P.falciparum): PF3D7_1357100; Gene product: elongation factor 1-alpha (eef1a)
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/d-type unit))
Phenotype Asexual bloodstage; Gametocyte/Gamete;
Last modified: 12 April 2011, 10:53
  *RMgm-28
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) : 16051702
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/ResearcherB. Franke-Fayard; C.J. Janse; A.P. Waters
Name Group/DepartmentLeiden Malaria Research Group
Name InstituteLeiden University Medical Center
CityLeiden
CountryThe Netherlands
Name of the mutant parasite
RMgm numberRMgm-28
Principal name222cl1
Alternative namePbGFP-LUCcon
Standardized name
Is the mutant parasite cloned after genetic modificationYes
Phenotype
Asexual blood stageGFP-Luciferase expression in all stages; increase of GFP-Luciferase expression during development from ring form into mature trophozoite/immature schizont; decrease of GFP-Luciferase expression in mature schizonts. Strong GFP-Luciferase expression in female gametocytes.
Gametocyte/GameteGFP-Luciferase expression in female and male gametocytes. Strong GFP-expression in female gametocytes. Higher expression in females, compared to males.
Fertilization and ookineteNot tested
OocystNot tested
SporozoiteNot tested
Liver stageNot tested
Additional remarks phenotype

Mutant/mutation
This mutant expresses GFP-luciferase under the control of the constitutive eef1a promoter.

Phenotype
GFP is expressed in all life cycle stages and expression of the transgene has no influence on parasite viability and infectivity.

Additional information
This mutant and mutant RMgm-333 have been used to analyse parasite distribution and parasite load in live animals (or in isolated organs) by real time in vivo imaging (by measurement of bioluminescence). In comparison with reporter mutants expressing only GFP under the same promoter (eef1a promoter), the GFP-fluorescence intensity of this reporter mutant is lower as a result of the GFP-Luciferase fusion.

Because all blood stages express luciferase, this mutant is less optimal for the analysis of the dynamics of schizont sequestration. Mutants are available that express GFP-Luciferase under the control of the schizont specific ama-1 promoter (RMgm-30RMgm-32). These mutants are more suitable for the analysis of schizont sequestration.
For analysis of parasite distribution and parasite load in tissues of live animals (or in isolated organs) by real time in vivo imaging the gametocyte-defective mutant RMgm-333 has the advantage over this mutant that imaging is not influenced by luciferase-expressing gametocytes that circulate in the peripheral blood and which express high levels of luciferase.

This mutant contains a drug-selectable marker (the pyrimethamine resistant tgdhfr gene). A reporter mutant (RMgm-29) expressing GFP-Luciferase under the control of the eefa promoter has been generated that lacks a drug selectable marker. This mutant which has been selected by FACS sorting, can be used for the introduction of additional genetic modifications using vectors that contain drug-selectable markers.

Other mutants
RMgm-333: A gametocyte-defective producing mutant expressing GFP-luciferase under the control of the constitutive eef1a promoter. This mutant has been generated in a non-gametocyte producer line of the ANKA strain of P. berghei (clone 2.33)


  Transgene: Mutant parasite expressing a transgene
Type and details of transgene
Is the transgene Plasmodium derived Transgene: not Plasmodium
Transgene nameA fusion of GFP (gfp-65) and Luciferase Firefly (LucIAV)
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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1 tatgcttgtc tcaaagatta agccatgcaa gtgaaagtat atgcacattt attgcagaaa ctgcgaacgg ctcattaaaa cagttataat ctacttgaca ttttattata aggataacta
121 cggaaaagct gtagctaata cttgttaagt acttttactc cccggagtaa ttgtatgtat ttgttaagac ccctaagaaa aaatgatatt aaaggaatta taacaaagaa gcaacacata
241 atataattat tcagtgtgta tcaatcgagt ttctgaccta tcagcttttg atgttagggt attgacctaa catggctttg acgggtaacg gggaattaga gttcgattcc ggagagggag
361 cctgagaaat agctaccaca tctaaggaag gcagcaggcg cgtaaattac ccaattctaa ataagagagg tagtgacaag aaataacaat ataaggccaa attttggttt tataattgga
481 atgatgggaa tttaaaacct tcccaaaaat caattggagg gcaagtctgg tgccagcagc cgcggtaatt ccagctccaa tagcgtatat taaaattgtt gcagttaaaa cgctcgtagt
601 tgaacttcaa gggtataatt attttaagca actcacttgg aaagaatcat gacttctgtc actgctttta tccttgttgc agttctttta atacagggcc ctttgagagc ccattaattt
721 atgactgggt ttctcgttac tttgagtaaa ttagagtgtt taaagcaaac agataaagcg tattttactg tgtttgaata ctatagcatg gaataacaac attgaatagg tcaaaagttt
841 ttgaaaaatt tttcttattt tggcttagat acagttaata ggagtagctt gggggcattt gtattcagat gtcagaggtg aaattcttag attttctgga gacaaacaac tgcgaaagca
961 tttgcctaaa atacttccat taatcaagaa cgaaagttaa gggagtgaag acgatcagat accgtcgtaa tcttaaccat aaactatgcc gactaagtgt tggatgaaaa tttataaata
1081 aaactatctt ctttaaagga gtagtttttt agatgcttcc ttcagtacct tatgagaaat caaagtcttt gggttctggg gcgagtattc gcgcaagcga gaaagttaaa agaattgacg
1201 gaagggcacc accaggcgtg gagcttgcgg cttaatttga ctcaacacgg ggaaactcac tagtttaaga caagagtagg attgacagat taatagctct ttcttgattt cttggatggt
1321 gatgcatggc cgtttttagt tcgtgaatat gatttgtctg gttaattccg ataacgaacg agatcttaac ctgctaatta gcggtggata tgtgatattc ttcgaaggtg gactaactat
1441 agcgttttcg aaggtatgtt gcataatcaa attggtttac cctttgtttt tttgtagcat attcttttat ttcgttgggt tttttcccta gtaaggatgt atctgcttta tttaatgctt
1561 cttagaggaa cgatgtgtgt ctaacacaag gaagtttaag gcaacaacag gtctgtgatg tccttagata tactaggctg cacgcgtgct acactgatat gtaaaacgag tatttaaaat
1681 tatatctgta tggtagataa tttaatttct acgtattatc agcatatact tttcctacac tgaaatagtg aaggtaatct ttatcaatac atatcgtgat ggggatagat tattgcaatt
1801 attaatcttg aacgaggaat gcctagtaag catgattcat cagattgtgc tgactacgtc cctgcccttt gtacacaccg cccgtcgctc ctaccgattg aaagatatga tgaattgttt
1921 ggacaagaaa atagaaattt tatttttatt ttttttggaa ggaccgtaaa tcctatcttt taaaggaagg agaagtcgta acaaggtttc cgtaggtgaa ttcactggcc gtcgttttac
2041 aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa tcgccttgca gcacatcccc ctttcgccag ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc caacagttgc
2161 gcagcctgaa tggcgaatgg cgcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg catatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc
2281 agccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg accgtctccg ggagctgcat gtgtcagagg ttttcaccgt
2401 catcaccgaa acgcgcgaga cgaaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat aatggtttct tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa
2521 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg
2641 tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg
2761 atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc
2881 aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga
3001 gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga
3121 atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact
3241 ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg
3361 ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac
3481 tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt
3601 tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
3721 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
3841 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
3961 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag cattgagaaa gcgccacgct tcccgaaggg agaaaggcgg
4081 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat
4201 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg
4321 attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc
4441 ctctccccgc gcgttggccg attcattaat gcagctggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaatg tgagttagct cactcattag gcaccccagg
4561 ctttacactt tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc acacaggaaa cagctatgac catgattacg ccaagcttgc atgcctgcag gtcaacaata
4681 aataataaat aaatattgtg gaaataaaat aacatataat tatttttaat acattgattt cccttttatt tttttaaatt tcattgatat aaaaatatat aataataaca tatatgattt
4801 caaattaatc ttttcaaaaa tggtgtttat ttttgtatgt ttgtgtatga attaatcaca taacacatct attaaattga gttggtaata tagacacaaa taaatatata tatttttata
4921 gcttaaaagt gtgttatgaa tattttaagc atattttctt tttctttgga ttgtgtaaaa tgaactcata taatgcgttt ttttgttttt gttattttgt cattttgtta ttttgctatt
5041 ttatggatta atttttgttt ataaaatggg aaataatttt aacatattta aataaatgga gaaaaaatat aaaataatta taaaaaaaag ttaatacaca ttttttcctg ttatagacct
5161 tatatttatt tatccatata tatatatata tatatatata tatatatata cataccaagt gaattaagag gaaagctaat ttattattca gaataatata tgaactatat ataattttta
5281 ttattttggt gtatattaat ctgtctatat gcatacatgc aataatttat cgacttatat atcaaataac ataaaataga agtgttttaa attatggata tatgctcaat attcattttt
5401 tttaataagt tagctatatt taaattatac attttatata tggtctcttt tttttttaaa tattatttaa gtgatcatga aaatataaat aatttttttt tatttaatat ccttttgctt
5521 gcatgtggta aatggaaatt tggatgtgtt ttgaargttc ggatatagtt gtatggacat ataatatatt ttgtgaaaaa ttggttttat gtttatactt atgccaatac tttttgagta
5641 aaacaaagca agtgcttata aataattaaa gccaatttta taatatatat ttttttattt aatttgaatt tagtagtata attttttatg gtaagtgctc aaagagagtt gcttataaag
5761 tatggtttgt ttctttttcg ccattttgaa ttacacatta aaaatatata gatacatata ttataatatg aaatcattaa taatttaggg aaattctaca aatttaaaaa cgaataaaat
5881 aattgttttt catcatgcca taacacaata ttgatatata catgtacaaa catttttttt atttggaaaa tataaattat ataaaaaaaa atgtatagta tacaaaatga gcatattcac
6001 acggggtgga cgttcatttt ttcatttttc ccctgttttt tatgagtata tgataaaatt ttatgaacat ttacacaaaa tgaaaatgga tatataggaa aaatggagcg gtatttcatt
6121 tatctttgat tgtcatttgg atattatatt accytgggta ggcaattaaa aatgttaaat aacaatttaa ggaaattata ttttatatat taaaattaac actgtattat atgattcgct
6241 tataaaagcc actctttccc catgcaaagc tgtttaatat caattttaac aaattacaca catgttaata tatttatata tataatttat atatttataa tttatatatt tatattttta
6361 ttatttatat atttattatt tattgtgtgt gtcaattcgg gtaggatata cctctttttt attgtttaaa gcgatttgta ttctaaaata taaagrattt gaaaaagaga aagatagaat
6481 atgatcccat catatatagc cctataattt ttatttagca gcgaattaat ttttctatta agtttatgtg taattaaaat aacggaatat atataataca ataaaaaagt gcataaatta
6601 aaattttttc aattaaattt ttttttttaa ggggttatat aatattaaat atataaaata cgattatata tttttgctac aattttttat attaagatat aaatagtaaa taaatggtat
6721 tatatggcat gtaatatata aattttttcc aatttttatt ttatatacac ttttcctttt tttgtcataa aacttaaaca atttacacat tcattttaaa aattgactat ttgtttcaac
6841 attttttgag tttccgtttt ataatagtat tttcatttgt atattgctta tatatataaa tacacaccta aatgttacaa aggatcaatg cataaaccgg tgtgtctggt cgtcgcgatg
6961 acccccaaga ggggcatcgg catcaacaac ggcctcccgt ggccccactt gaccacagat ttcaaacact tttctcgtgt gacaaaaacg acgcccgaag aagccagtcg cctgaacggg
7081 tggcttccca ggaaatttgc aaagacgggc gactctggac ttccctctcc atcagtcggc aagagattca acgccgttgt catgggacgg aaaacctggg aaagcatgcc tcgaaagttt
7201 agacccctcg tggacagatt gaacatcgtc gtttcctctt ccctcaaaga agaagacatt gcggcggaga agcctcaagc tgaaggccag cagcgcgtcc gagtctgtgc ttcactccca
7321 gcagctctca gccttctgga ggaagagtac aaggattctg tcgaccagat ttttgtcgtg ggaggagcgg gactgtacga ggcagcgctg tctctgggcg ttgcctctca cctgtacatc
7441 acgcgtgtag cccgcgagtt tccgtgcgac gttttcttcc ctgcgttccc cggagatgac attctttcaa acaaatcaac tgctgcgcag gctgcagctc ctgccgagtc tgtgttcgtt
7561 cccttttgtc cggagctcgg aagagagaag gacaatgaag cgacgtatcg acccatcttc atttccaaga ccttctcaga caacggggtt ccctacgact ttgtggttct cgagaagaga
7681 aggaagactg acgacgcagc cactgcggaa ccgagcaacg caatgagctc cttgacgtcc acgagggaga caactcccgt gcacgggttg caggctcctt cttcggccgc agccattgcc
7801 ccggtgttgg cgtggatgga cgaagaagac cggaaaaaac gcgagcaaaa ggaactgatt cgggccgttc cgcatgttca ctttagaggc catgaagagt tccagtacct tgatctcatt
7921 gccgacatta ttaacaatgg aaggacaatg gatgaccgaa cgggcgttgg tgtcatctcc aaattcggct gcactatgcg ctactcgctg gatcaggcct ttccacttct caccacaaag
8041 cgtgtgttct ggaaaggggt cctcgaagag ttgctgtggt tcattcgcgg cgacacgaac gcaaaccatc tttctgagaa gggcgtgaag atctgggaca agaatgtgac acgcgagttc
8161 ctcgattcgc gcaatctccc ccaccgagag gtcggagaca tcggcccggg ctacggcttc cagtggagac acttcggcgc ggcatacaaa gacatgcaca cagactacac agggcagggc
8281 gtcgaccagc tgaagaatgt gatccagatg ctgagaacga atccaacaga tcgtcgcatg ctcatgactg cctggaatcc tgcagcgctg gacgaaatgg cgctgccgcc ttgtcacttg
8401 ttgtgccagt tctacgtgaa cgaccagaag gagctgtcgt gcatcatgta tcagcggtcg tgcgatgtcg gcctcggcgt ccccttcaac atcgcttcct attcgctttt gacgctcatg
8521 gttgcacacg tctgcaacct aaaacctaag gagttcattc acttcatggg gaacacgcat gtctacacga accatgtcga ggctttaaaa gagcagctgc ggagagaacc gagaccgttc
8641 cccattgtga acatcctcaa caaggaacgc atcaaggaaa tcgacgattt caccgccgag gattttgagg tcgtgggcta cgtcccgcac ggacgaatcc agatggagat ggctgtctag
8761 cggaaataca gaagctagct ttgatcccgt ttttcttact tatatattta taccaattga ttgtatttat aactgtaaaa atgtgtatgt tgtgtgcata tttttttttg tgcatgcaca
8881 tgcatgtaaa tagctaaaat tatgaacatt ttattttttg ttcagaaaaa aaaaacttta cacacataaa atggctagta tgaatagcca tattttatat aaattaaatc ctatgaattt
9001 atgaccatat taaaaattta gatatttatg gaacataata tgtttgaaac aataagacaa aattattatt attattatta tttttactgt tataattatg ttgtctcttc aatgattcat
9121 aaatagttgg acttgatttt taaaatgttt ataatatgat tagcatagtt aaataaaaaa agttgaaaaa ttaaaaaaaa acatataaac acaaatgatg ttttttcctt caatttcgat
9241 tgataattac tgcagcccag cttaattctt ttcgagctct ttatgcttaa gtttacaatt taatattcat actttaagta ttttttgtag tatcctagat attgtgcttt aaatgctcac
9361 ccctcaaagc accagtaata ttttcatcca ctgaaatacc attaaatttt caaaaaaata ctatgcatat aatgttatac atataaacat aaaacgccat gtaaatcaaa aaatatataa
9481 aaatatgtat aaaaataaat atgcactaaa tataagctaa ttatgcataa aaattaaagt gccctttatt aactagtcgt aattatttat atttctatgt tataaaaaaa tcctcatata
9601 ataatataat taatatatgt aatgtttttt ttattttata attttaatat aaaataatat gtaaattaat tcaaaaaata aatataattg ttgtgaaaca aaaaacgtaa ttttttcatt
9721 tgccttcaaa atttaaattt attttaatat ttcctaaaat atatatactt tgtgtataaa tatataaaaa tatatatttg cttataaata aataaaaaat tttataaaac atagggggat
9841 ctatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc gatgccacct
9961 acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac caccttcacc tacggcgtgc agtgcttcag ccgctacccc gaccacatga
10081 agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc
10201 tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat catggccgac aagcagaaga
10321 acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg cccgacaacc
10441 actacctgag cacccagtcc gccctgagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactcacgg catggacgag ctgtacccgg
10561 ggatcctggc tagccagtcg acctgcaggc atgcaagctt gcggccgatc caaatggaag acgccaaaaa cataaagaaa ggcccggcgc cattctatcc gctggaagat ggaaccgctg
10681 gagagcaact gcataaggct atgaagagat acgccctggt tcctggaaca attgctttta cagatgcaca tatcgaggtg aacatcacgt acgcggaata cttcgaaatg tccgttcggt
10801 tggcagaagc tatgaaacga tatgggctga atacaaatca cagaatcgtc gtatgcagtg aaaactctct tcaattcttt atgccggtgt tgggcgcgtt atttatcgga gttgcagttg
10921 cgcccgcgaa cgacatttat aatgaacgtg aattgctcaa cagtatgaac atttcgcagc ctaccgtagt gtttgtttcc aaaaaggggt tgcaaaaaat tttgaacgtg caaaaaaaat
11041 taccaataat ccagaaaatt attatcatgg attctaaaac ggattaccag ggatttcagt cgatgtacac gttcgtcaca tctcatctac ctcccggttt taatgaatac gattttgtac
11161 cagagtcctt tgatcgtgac aaaacaattg cactgataat gaattcctct ggatctactg ggttacctaa gggtgtggcc cttccgcata gaactgcctg cgtcagattc tcgcatgcca
11281 gagatcctat ttttggcaat caaatcattc cggatactgc gattttaagt gttgttccat tccatcacgg ttttggaatg tttactacac tcggatattt gatatgtgga tttcgagtcg
11401 tcttaatgta tagatttgaa gaagagctgt ttttacgatc ccttcaggat tacaaaattc aaagtgcgtt gctagtacca accctatttt cattcttcgc caaaagcact ctgattgaca
11521 aatacgattt atctaattta cacgaaattg cttctggggg cgcacctctt tcgaaagaag tcggggaagc ggttgcaaaa cgcttccatc ttccagggat acgacaagga tatgggctca
11641 ctgagactac atcagctatt ctgattacac ccgaggggga tgataaaccg ggcgcggtcg gtaaagttgt tccatttttt gaagcgaagg ttgtggatct ggataccggg aaaacgctgg
11761 gcgttaatca gagaggcgaa ttatgtgtca gaggacctat gattatgtcc ggttatgtaa acaatccgga agcgaccaac gccttgattg acaaggatgg atggctacat tctggagaca
11881 tagcttactg ggacgaagac gaacacttct tcatagttga ccgcttgaag tctttaatta aatacaaagg atatcaggtg gcccccgctg aattggaatc gatattgtta caacacccca
12001 acatcttcga cgcgggcgtg gcaggtcttc ccgacgatga cgccggtgaa cttcccgccg ccgttgttgt tttggagcac ggaaagacga tgacggaaaa agagatcgtg gattacgtcg
12121 ccagtcaagt aacaaccgcg aaaaagttgc gcggaggagt tgtgtttgtg gacgaagtac cgaaaggtct taccggaaaa ctcgacgcaa gaaaaatcag agagatcctc ataaaggcca
12241 agaagggcgg aaagatcgcc gtgtaattct agaagatccc gtttttctta cttatatatt tataccaatt gattgtattt ataactgtaa aaatgtgtat gttgtgtgca tatttttttt
12361 tgtgcatgca catgcatgta aatagctaaa attatgaaca ttttattttt tgttcagaaa aaaaaaactt tacacacata aaatggctag tatgaatagc catattttat ataaattaaa
12481 tcctatgaat ttatgaccat attaaaaatt tagatattta tggaacataa tatgtttgaa acaataagac aaaattatta ttattattat tatttttact gttataatta tgttgtctct
12601 tcaatgattc ataaatagtt ggacttgatt tttaaaatgt ttataatatg attagcatag ttaaataaaa aaagttgaaa aattaaaaaa aaacatataa acacaaatga tgttttttcc
12721 ttcaatttcg ggtaccgagc tcgaattcaa cctggttgat cttgccagta gtca
Restriction sites to linearize plasmid ApaI SacII
Selectable marker used to select the mutant parasitetgdhfr
Promoter of the selectable markerpbdhfr
Selection (positive) procedurepyrimethamine
Selection (negative) procedureNo
Additional remarks genetic modificationThe GFP-Luciferase 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. In this mutant multiple copies (2-3) of the GFP-Luciferase gene are integrated into the small subunit ribosomal rna gene (c/d-type unit). No gene model is yet available for this integration locus!
Additional remarks selection procedure
Other details transgene
Promoter
Gene Model of Parasite PBANKA_1133300
Gene Model P. falciparum ortholog PF3D7_1357100
Gene productelongation factor 1-alpha
Gene product: Alternative nameeef1a
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
Additional information primer 1
Sequence Primer 2
Additional information primer 2
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/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