Summary

RMgm-30
Malaria parasiteP. berghei
Genotype
Transgene
Transgene not Plasmodium: A fusion of GFP (gfp-mu3) and Luciferase Firefly (LucIAV)
Promoter: Gene model: PBANKA_0915000; Gene model (P.falciparum): PF3D7_1133400; Gene product: apical membrane antigen 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 (d-type unit))
Phenotype Asexual bloodstage;
Last modified: 12 April 2011, 09:38
  *RMgm-30
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 MRA-866
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-30
Principal name354cl4
Alternative namePbGFP-LUCsch
Standardized name
Is the mutant parasite cloned after genetic modificationYes
Phenotype
Asexual blood stageGFP-Luciferase expression in mature schizonts and young ring forms.
Gametocyte/GameteNot different from wild type
Fertilization and ookineteNot tested
OocystNot tested
SporozoiteNot tested
Liver stageNot tested
Additional remarks phenotype

Mutant/mutation
The mutant expresses GFP-luciferase under the control of the schizont-specific ama-1 promoter.

Phenotype
GFP-Luciferase is expressed in maturing schizonts and is still present in young ringforms (0-4 hours after invasion), carried over from the schizonts. No expression of GFP-luciferase in gametocytes/gametes

Additional information
This mutant has been used to analyse CD36-mediated sequestration of the schizont stage of P. berghei in live animals (or in isolated organs) by real time in vivo imaging (by measurement of bioluminescence) (Franke-Fayard et al., 2005, Proc Natl Sci USA, 102, 11468-73). These studies indicate that the schizont of P. berghei mainly sequester/accumulate in small capillaries of adipose tissue, lungs and the spleen.

In comparison with mutants expressing only GFP under the same promoter (ama1 promoter), the GFP-fluorescence intensity of this mutant is lower as a result of the GFP-Luciferase fusion.

This mutant contains a drug-selectable marker (the pyrimethamine resistant tgdhfr gene). A  mutant (RMgm-32) expressing GFP-Luciferase under the control of the ama-1 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.


  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-mu3) 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 atcgaattcc catggaacca aaataacata ttaatgaatt ttcttttatt gtgaaaatgt tcttatttgt tttaaaaaat caaagtaaat ttaattcctt taattgtgct aaatattgtg
9361 tttgtataac tataatgaca tttcctatgc ttttttaata ttatatataa ttttttgtgt tttttcaaaa aaaacgtgta tttgttttta tttgagtgta caatttgcat agtgagttta
9481 aagtgtttga aaattattta atttataatt ttcataaaac gtttaatcat ttatcaaaat tgcggttgca atatagacag aaaatattaa cttaattttc attgttttgt aaaacatttt
9601 cattattttt tttaacaaaa ttaaagtaaa tgagatggta aattatttta attataacca tttgtttaat tttttttatt ttattactat tatttttttt cttttgtgct gcgcaattaa
9721 aaagagaatt aagggtgcac caatatattt aagcaatctt ttgcgacaca caagaaaaaa tataacatca acatatttaa aataatggaa attttttaat aatttattct atatatattt
9841 aatatatgta aaaaaactaa tattgtaata taattaaata aaacttgaaa ttgatataat atgtatatat attgcatgtt tagctttaaa taatatttaa gtattgattt tctttacaca
9961 caaaaacatt tgccttttta tgtgaggatt attgtttctt tatttatatt gctatgaatt tattcttagt ataaatttat tttttttcgg ctttgaaaat aattttatat atataaaaag
10081 aaaaaatatg gaaaataaaa ttcataaatc atatttgaca catttcaatt tttttttctt atttcacagc tagccatata ataatatata tgtgtctatg ggttcaaata gcttaatttc
10201 aatgtgcgtg attttttttt ataaaaaaac taattttatg tttggttatg catgcaactg ttataaaata aaaattgtaa aatttgtttt tttgtacaca tatttgtact ttttttaatg
10321 cacatgaaaa aagaaaggaa aaaaaagggg gaaaaaaaag gaaaaaaaaa gggaaaaaag ggaaaaaacg tacatctacg cattgttatt tagcaataaa atagagaaca aatctataaa
10441 tatataatgt gtacaacaaa ttaatttaat ttatattttt actgtaattt cataataatt ttcattttta cattttaatg tttttttaat tggaatataa ttcaaaatga ttttgacaca
10561 ttttacttat agaagaaggc tatataatta tataagtgtg tatatataag tattgtatgg taattgtttt ataaattaaa tttttaaaaa acattattct attttataat ttgtaaatta
10681 aaaatattaa taaaataaaa cgatataaaa ggatctatga gtaaaggaga agaacttttc actggagttg tcccaattct tgttgaatta gatggtgatg ttaatgggca caaattttct
10801 gtcagtggag agggtgaagg tgatgcaaca tacggaaaac ttacccttaa atttatttgc actactggaa aactacctgt tccatggcca acacttgtca ctactttcgg ttatggtgtt
10921 caatgctttg cgagataccc agatcatatg aaacagcatg actttttcaa gagtgccatg cccgaaggtt atgtacagga aagaactata tttttcaaag atgacgggaa ctacaagaca
11041 cgtgctgaag tcaagtttga aggtgatacc cttgttaata gaatcgagtt aaaaggtatt gattttaaag aagatggaaa cattcttgga cacaaattgg aatacaacta taactcacac
11161 aatgtataca tcatggcaga caaacaaaag aatggaatca aagttaactt caaaattaga cacaacattg aagatggaag cgttcaacta gcagaccatt atcaacaaaa tactccaatt
11281 ggcgatggcc ctgtcctttt accagacaac cattacctgt ccacacaatc tgccctttcg aaagatccca acgaaaagag agaccacatg gtccttcttg agtttgtaac agctgctggg
11401 attacacatg gcatggatga actatacaaa gggatcctgg ctagccagtc gacctgcagg catgcaagct tgcggccgat ccaaatggaa gacgccaaaa acataaagaa aggcccggcg
11521 ccattctatc cgctggaaga tggaaccgct ggagagcaac tgcataaggc tatgaagaga tacgccctgg ttcctggaac aattgctttt acagatgcac atatcgaggt gaacatcacg
11641 tacgcggaat acttcgaaat gtccgttcgg ttggcagaag ctatgaaacg atatgggctg aatacaaatc acagaatcgt cgtatgcagt gaaaactctc ttcaattctt tatgccggtg
11761 ttgggcgcgt tatttatcgg agttgcagtt gcgcccgcga acgacattta taatgaacgt gaattgctca acagtatgaa catttcgcag cctaccgtag tgtttgtttc caaaaagggg
11881 ttgcaaaaaa ttttgaacgt gcaaaaaaaa ttaccaataa tccagaaaat tattatcatg gattctaaaa cggattacca gggatttcag tcgatgtaca cgttcgtcac atctcatcta
12001 cctcccggtt ttaatgaata cgattttgta ccagagtcct ttgatcgtga caaaacaatt gcactgataa tgaattcctc tggatctact gggttaccta agggtgtggc ccttccgcat
12121 agaactgcct gcgtcagatt ctcgcatgcc agagatccta tttttggcaa tcaaatcatt ccggatactg cgattttaag tgttgttcca ttccatcacg gttttggaat gtttactaca
12241 ctcggatatt tgatatgtgg atttcgagtc gtcttaatgt atagatttga agaagagctg tttttacgat cccttcagga ttacaaaatt caaagtgcgt tgctagtacc aaccctattt
12361 tcattcttcg ccaaaagcac tctgattgac aaatacgatt tatctaattt acacgaaatt gcttctgggg gcgcacctct ttcgaaagaa gtcggggaag cggttgcaaa acgcttccat
12481 cttccaggga tacgacaagg atatgggctc actgagacta catcagctat tctgattaca cccgaggggg atgataaacc gggcgcggtc ggtaaagttg ttccattttt tgaagcgaag
12601 gttgtggatc tggataccgg gaaaacgctg ggcgttaatc agagaggcga attatgtgtc agaggaccta tgattatgtc cggttatgta aacaatccgg aagcgaccaa cgccttgatt
12721 gacaaggatg gatggctaca ttctggagac atagcttact gggacgaaga cgaacacttc ttcatagttg accgcttgaa gtctttaatt aaatacaaag gatatcaggt ggcccccgct
12841 gaattggaat cgatattgtt acaacacccc aacatcttcg acgcgggcgt ggcaggtctt cccgacgatg acgccggtga acttcccgcc gccgttgttg ttttggagca cggaaagacg
12961 atgacggaaa aagagatcgt ggattacgtc gccagtcaag taacaaccgc gaaaaagttg cgcggaggag ttgtgtttgt ggacgaagta ccgaaaggtc ttaccggaaa actcgacgca
13081 agaaaaatca gagagatcct cataaaggcc aagaagggcg gaaagatcgc cgtgtaattc tagaagatcc cgtttttctt acttatatat ttataccaat tgattgtatt tataactgta
13201 aaaatgtgta tgttgtgtgc atattttttt ttgtgcatgc acatgcatgt aaatagctaa aattatgaac attttatttt ttgttcagaa aaaaaaaact ttacacacat aaaatggcta
13321 gtatgaatag ccatatttta tataaattaa atcctatgaa tttatgacca tattaaaaat ttagatattt atggaacata atatgtttga aacaataaga caaaattatt attattatta
13441 ttatttttac tgttataatt atgttgtctc ttcaatgatt cataaatagt tggacttgat ttttaaaatg tttataatat gattagcata gttaaataaa aaaagttgaa aaattaaaaa
13561 aaaacatata aacacaaatg atgttttttc cttcaatttc gggtaccgag ctcgaattca acctggttga tcttgccagt agtca
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 (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_0915000
Gene Model P. falciparum ortholog PF3D7_1133400
Gene productapical membrane antigen 1
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
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 (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