Summary

RMgm-86
Malaria parasiteP. berghei
Genotype
Transgene
Transgene not Plasmodium: drFP583/DsRed/RFP, RedStar
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: PBANKA_0306000; Gene product: 6-cysteine protein (230p)
Phenotype Asexual bloodstage; Gametocyte/Gamete; Fertilization and ookinete; Oocyst; Sporozoite; Liver stage;
Last modified: 26 June 2009, 16:01
  *RMgm-86
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) : 19026596
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 line
The mutant parasite was generated by
Name PI/ResearcherB. Franke-Fayard; C.J. Janse
Name Group/DepartmentLeiden Malaria Research Group
Name InstituteLeiden University Medical Center
CityLeiden
CountryThe Netherlands
Name of the mutant parasite
RMgm numberRMgm-86
Principal name733cl1
Alternative nameP. berghei-RFP; P. berghei-RedStar
Standardized name
Is the mutant parasite cloned after genetic modificationYes
Phenotype
Asexual blood stageStrong RFP (RedStar) expression in all stages except for male gametes; Increase of RFP expression during development from ring form into mature trophozoite/immature schizont; decrease of RFP expression in mature schizonts. Strong RFP-expression in female gametocytes.
Gametocyte/GameteRFP (RedStar) expression in female and male gametocytes. Strong RFP-expression in female gametocytes. Higher expression in females, compared to males.
Fertilization and ookineteRFP (RedStar) expression in ookinetes.
OocystRFP (RedStar) expression in oocysts.
SporozoiteRFP (RedStar) expression in sporozoites.
Liver stageRFP (RedStar) expression during liver stage development.
Additional remarks phenotype

Mutant/mutation
The mutant expresses RFP (RedStar) under the control of the constitutive promoter of the elongation factor 1 alpha (eef1a) gene of P. berghei.

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

Additional information
This mutant has been used to visualise and analyse liver stage development (Sturm, A. et al., (2008). Protist, Nov. 19.

See also mutant lines RMgm-295 and RMgm-296 expressing the red fluorescent proteins tdTomato and mCherry  under the control of the eef1a promoter.


  Transgene: Mutant parasite expressing a transgene
Type and details of transgene
Is the transgene Plasmodium derived Transgene: not Plasmodium
Transgene namedrFP583/DsRed/RFP, RedStar
Details of the genetic modification
Inducable system usedNo
Additional remarks inducable system
Type of plasmid/constructPlasmid double 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 aattcactgg ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt tacccaactt
61 aatcgccttg cagcacatcc ccctttcgcc agctggcgta atagcgaaga ggcccgcacc
121 gatcgccctt cccaacagtt gcgcagcctg aatggcgaat ggcgcctgat gcggtatttt
181 ctccttacgc atctgtgcgg tatttcacac cgcatatggt gcactctcag tacaatctgc
241 tctgatgccg catagttaag ccagccccga cacccgccaa cacccgctga cgcgccctga
301 cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc
361 atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga gacgaaaggg cctcgtgata
421 cgcctatttt tataggttaa tgtcatgata ataatggttt cttagacgtc aggtggcact
481 tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg
541 tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt
601 atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct
661 gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca
721 cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc
781 gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc
841 cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg
901 gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta
961 tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc
1021 ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt
1081 gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg
1141 cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct
1201 tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc
1261 tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct
1321 cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac
1381 acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc
1441 tcactgatta agcattggta actgtcagac caagtttact catatatact ttagattgat
1501 ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg
1561 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc
1621 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
1681 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
1741 gtaactggct tcagcagagc gcagatacca aatactgtcc ttctagtgta gccgtagtta
1801 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
1861 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
1921 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
1981 gagcgaacga cctacaccga actgagatac ctacagcgtg agcattgaga aagcgccacg
2041 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
2101 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
2161 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
2221 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
2281 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct
2341 gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa
2401 gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg
2461 cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tgtgagttag
2521 ctcactcatt aggcacccca ggctttacac tttatgcttc cggctcgtat gttgtgtgga
2581 attgtgagcg gataacaatt tcacacagga aacagctatg accatgatta cgccaagctt
2641 ccgcgggtat atggtaaaga acctactaac acaataaaat atttaaataa tgtatttcct
2701 ataaataaat ttacagattt attttttaat acaaaagata tagatatacc agaaataaat
2761 gatcagttta aaggttttaa attctttatg acatcattta taaatcatgg atcatatcca
2821 ctaacaatag aatgtggtgt aacaaatggt ggaactagtt ataaaagagc aattatttta
2881 ttgcatgttc gaactgattt aaaagataga ccagtttcat tttgtgattt tcgaaaagga
2941 gaattatata attatttgaa tgcttatact gaaggggatg tatgcataat aatttccaaa
3001 tcaaatacaa gttttggttt tagatgccca gtaaatacaa aaaaaatgcc aaaaaattgt
3061 tttacgcaag tatatgaaaa agggtatcta aatgacgcca ataaaattaa tactaaaaat
3121 gttattaact attcatttga aaatccagaa tatgcgctag ctggttytaa ttatacatta
3181 acaaaatcgt atcaatttga atgtcattgt gtagataaag aaacagaaca aattgtaaaa
3241 acggttttag tcaaatatgt aaatgaagat gaaatatatg attataatga ttttccaatg
3301 gtgaatcaca aacctattat tgcacatcca aataaaacac atcaagcttg catgcctgca
3361 ggtcaacaat aaataataaa taaatattgt ggaaataaaa taacatataa ttatttttaa
3421 tacattgatt tcccttttat ttttttaaat ttcattgata taaaaatata taataataac
3481 atatatgatt tcaaattaat cttttcaaaa atggtgttta tttttgtatg tttgtgtatg
3541 aattaatcac ataacacatc tattaaattg agttggtaat atagacacaa ataaatatat
3601 atatttttat agcttaaaag tgtgttatga atattttaag catattttct ttttctttgg
3661 attgtgtaaa atgaactcat ataatgcgtt tttttgtttt tgttattttg tcattttgtt
3721 attttgctat tttatggatt aatttttgtt tataaaatgg gaaataattt taacatattt
3781 aaataaatgg agaaaaaata taaaataatt ataaaaaaaa gttaatacac attttttcct
3841 gttatagacc ttatatttat ttatccatat atatatatat atatatatat atatatatat
3901 acataccaag tgaattaaga ggaaagctaa tttattattc agaataatat atgaactata
3961 tataattttt attattttgg tgtatattaa tctgtctata tgcatacatg caataattta
4021 tcgacttata tatcaaataa cataaaatag aagtgtttta aattatggat atatgctcaa
4081 tattcatttt ttttaataag ttagctatat ttaaattata cattttatat atggtctctt
4141 ttttttttaa atattattta agtgatcatg aaaatataaa taattttttt ttatttaata
4201 tccttttgct tgcatgtggt aaatggaaat ttggatgtgt tttgaargtt cggatatagt
4261 tgtatggaca tataatatat tttgtgaaaa attggtttta tgtttatact tatgccaata
4321 ctttttgagt aaaacaaagc aagtgcttat aaataattaa agccaatttt ataatatata
4381 tttttttatt taatttgaat ttagtagtat aattttttat ggtaagtgct caaagagagt
4441 tgcttataaa gtatggtttg tttctttttc gccattttga attacacatt aaaaatatat
4501 agatacatat attataatat gaaatcatta ataatttagg gaaattctac aaatttaaaa
4561 acgaataaaa taattgtttt tcatcatgcc ataacacaat attgatatat acatgtacaa
4621 acattttttt tatttggaaa atataaatta tataaaaaaa aatgtatagt atacaaaatg
4681 agcatattca cacggggtgg acgttcattt tttcattttt cccctgtttt ttatgagtat
4741 atgataaaat tttatgaaca tttacacaaa atgaaaatgg atatatagga aaaatggagc
4801 ggtatttcat ttatctttga ttgtcatttg gatattatat taccytgggt aggcaattaa
4861 aaatgttaaa taacaattta aggaaattat attttatata ttaaaattaa cactgtatta
4921 tatgattcgc ttataaaagc cactctttcc ccatgcaaag ctgtttaata tcaattttaa
4981 caaattacac acatgttaat atatttatat atataattta tatatttata atttatatat
5041 ttatattttt attatttata tatttattat ttattgtgtg tgtcaattcg ggtaggatat
5101 acctcttttt tattgtttaa agcgatttgt attctaaaat ataaagratt tgaaaaagag
5161 aaagatagaa tatgatccca tcatatatag ccctataatt tttatttagc agcgaattaa
5221 tttttctatt aagtttatgt gtaattaaaa taacggaata tatataatac aataaaaaag
5281 tgcataaatt aaaatttttt caattaaatt ttttttttta aggggttata taatattaaa
5341 tatataaaat acgattatat atttttgcta caatttttta tattaagata taaatagtaa
5401 ataaatggta ttatatggca tgtaatatat aaattttttc caatttttat tttatataca
5461 cttttccttt ttttgtcata aaacttaaac aatttacaca ttcattttaa aaattgacta
5521 tttgtttcaa cattttttga gtttccgttt tataatagta ttttcatttg tatattgctt
5581 atatatataa atacacacct aaatgttaca aaggatcaat gcataaaccg gtgtgtctgg
5641 tcgtcgcgat gacccccaag aggggcatcg gcatcaacaa cggcctcccg tggccccact
5701 tgaccacaga tttcaaacac ttttctcgtg tgacaaaaac gacgcccgaa gaagccagtc
5761 gcctgaacgg gtggcttccc aggaaatttg caaagacggg cgactctgga cttccctctc
5821 catcagtcgg caagagattc aacgccgttg tcatgggacg gaaaacctgg gaaagcatgc
5881 ctcgaaagtt tagacccctc gtggacagat tgaacatcgt cgtttcctct tccctcaaag
5941 aagaagacat tgcggcggag aagcctcaag ctgaaggcca gcagcgcgtc cgagtctgtg
6001 cttcactccc agcagctctc agccttctgg aggaagagta caaggattct gtcgaccaga
6061 tttttgtcgt gggaggagcg ggactgtacg aggcagcgct gtctctgggc gttgcctctc
6121 acctgtacat cacgcgtgta gcccgcgagt ttccgtgcga cgttttcttc cctgcgttcc
6181 ccggagatga cattctttca aacaaatcaa ctgctgcgca ggctgcagct cctgccgagt
6241 ctgtgttcgt tcccttttgt ccggagctcg gaagagagaa ggacaatgaa gcgacgtatc
6301 gacccatctt catttccaag accttctcag acaacggggt tccctacgac tttgtggttc
6361 tcgagaagag aaggaagact gacgacgcag ccactgcgga accgagcaac gcaatgagct
6421 ccttgacgtc cacgagggag acaactcccg tgcacgggtt gcaggctcct tcttcggccg
6481 cagccattgc cccggtgttg gcgtggatgg acgaagaaga ccggaaaaaa cgcgagcaaa
6541 aggaactgat tcgggccgtt ccgcatgttc actttagagg ccatgaagag ttccagtacc
6601 ttgatctcat tgccgacatt attaacaatg gaaggacaat ggatgaccga acgggcgttg
6661 gtgtcatctc caaattcggc tgcactatgc gctactcgct ggatcaggcc tttccacttc
6721 tcaccacaaa gcgtgtgttc tggaaagggg tcctcgaaga gttgctgtgg ttcattcgcg
6781 gcgacacgaa cgcaaaccat ctttctgaga agggcgtgaa gatctgggac aagaatgtga
6841 cacgcgagtt cctcgattcg cgcaatctcc cccaccgaga ggtcggagac atcggcccgg
6901 gctacggctt ccagtggaga cacttcggcg cggcatacaa agacatgcac acagactaca
6961 cagggcaggg cgtcgaccag ctgaagaatg tgatccagat gctgagaacg aatccaacag
7021 atcgtcgcat gctcatgact gcctggaatc ctgcagcgct ggacgaaatg gcgctgccgc
7081 cttgtcactt gttgtgccag ttctacgtga acgaccagaa ggagctgtcg tgcatcatgt
7141 atcagcggtc gtgcgatgtc ggcctcggcg tccccttcaa catcgcttcc tattcgcttt
7201 tgacgctcat ggttgcacac gtctgcaacc taaaacctaa ggagttcatt cacttcatgg
7261 ggaacacgca tgtctacacg aaccatgtcg aggctttaaa agagcagctg cggagagaac
7321 cgagaccgtt ccccattgtg aacatcctca acaaggaacg catcaaggaa atcgacgatt
7381 tcaccgccga ggattttgag gtcgtgggct acgtcccgca cggacgaatc cagatggaga
7441 tggctgtcta ccggaaatac agaagctagc tttgatcccg tttttcttac ttatatattt
7501 ataccaattg attgtattta taactgtaaa aatgtgtatg ttgtgtgcat attttttttt
7561 gtgcatgcac atgcatgtaa atagctaaaa ttatgaacat tttatttttt gttcagaaaa
7621 aaaaaacttt acacacataa aatggctagt atgaatagcc atattttata taaattaaat
7681 cctatgaatt tatgaccata ttaaaaattt agatatttat ggaacataat atgtttgaaa
7741 caataagaca aaattattat tattattatt atttttactg ttataattat gttgtctctt
7801 caatgattca taaatagttg gacttgattt ttaaaatgtt tataatatga ttagcatagt
7861 taaataaaaa aagttgaaaa attaaaaaaa aacatataaa cacaaatgat gttttttcct
7921 tcaatttcga ttgataatta ctgcagccca gcttaattct tttcgagctc tttatgctta
7981 agtttacaat ttaatattca tactttaagt attttttgta gtatcctaga tattgtgctt
8041 taaatgctca cccctcaaag caccagtaat attttcatcc actgaaatac cattaaattt
8101 tcaaaaaaat actatgcata taatgttata catataaaca taaaacgcca tgtaaatcaa
8161 aaaatatata aaaatatgta taaaaataaa tatgcactaa atataagcta attatgcata
8221 aaaattaaag tgccctttat taactagtcg taattattta tatttctatg ttataaaaaa
8281 atcctcatat aataatataa ttaatatatg taatgttttt tttattttat aattttaata
8341 taaaataata tgtaaattaa ttcaaaaaat aaatataatt gttgtgaaac aaaaaacgta
8401 attttttcat ttgccttcaa aatttaaatt tattttaata tttcctaaaa tatatatact
8461 ttgtgtataa atatataaaa atatatattt gcttataaat aaataaaaaa ttttataaaa
8521 cataggggga tccaaaatga gtagatcttc taagaacgtc atcaaggaat tcatgagatt
8581 caaggttaaa atggaaggta ctgttaacgg ccacgaattc gaaatcgaag gtgaaggtga
8641 gggtagacca tatgaaggtc acaacacagt caagttgaag gttactaagg gtggtccact
8701 gccattcgct tgggacatct tgtctccaca attccaatac ggttctaagg tctacgtcaa
8761 gcacccagct gacattccag actacaagaa gttgtccttc ccagaaggtt tcaagtggga
8821 aaggatcatg aacttcgaag acggtggcgt tgttactgtt actcaagact cctccttgca
8881 agacggttgt ttcatctaca aggtcaagct cattggtgtc aacttcccat ctgacggtcc
8941 agtcatgcaa aagaagacta tgggttggga agcttctacc gaacgtttgt acccaagaga
9001 cggtgtcttg aagggtgaaa tccacaaggc cttgaagttg aaggacggtg gtcactactt
9061 ggtcgaattc aagtctatct acaaggccaa gaagcaagtc caattgccag gctattacta
9121 cgttgactct aagttggaca tcatctctca caacgaagac tacactatcg tcgaacaata
9181 cgaacgtact gaaggtagac accacttgtt cttgtaaact agttctagaa gatcccgttt
9241 ttcttactta tatatttata ccaattgatt gtatttataa ctgtaaaaat gtgtatgttg
9301 tgtgcatatt tttttttgtg catgcacatg catgtaaata gctaaaatta tgaacatttt
9361 attttttgtt cagaaaaaaa aaactttaca cacataaaat ggctagtatg aatagccata
9421 ttttatataa attaaatcct atgaatttat gaccatatta aaaatttaga tatttatgga
9481 acataatatg tttgaaacaa taagacaaaa ttattattat tattattatt tttactgtta
9541 taattatgtt gtctcttcaa tgattcataa atagttggac ttgattttta aaatgtttat
9601 aatatgatta gcatagttaa ataaaaaaag ttgaaaaatt aaaaaaaaac atataaacac
9661 aaatgatgtt ttttccttca atttcgggta ccgagctcga attctcttga gcccgttaat
9721 gaaatagata caattcattc atgttatata catctagaac ataatctgaa tatggttcaa
9781 gttaaatgtc caaaaattat aaaaagtgat gatatttttg atggtaatac cataatagac
9841 accaaggtaa catcacgaag tagtcaacaa aataattttt atttagaaaa tacagatgtt
9901 gaaccagaag aaatagagaa atataaaaat atagaataca taccagaaaa cgatgaagta
9961 atgcatctag acaaaaaaga aaagctagat gatatattac caggtgttat catattagat
10021 aaaaataaaa tgttcaaaga aaaaggacat ttcacttttg ttactccatt aattgtagaa
10081 aaggtattaa tattaaaaat atattgtgat aatactaaaa caataattaa taatatgaaa
10141 gggaaaaaag gtattacagt aataaggatt tctcaaaata caacaaaaaa taaattttat
10201 ggatgtgact tttcaggtaa ttctaaaaaa acattttact attccaatgt ttatgattta
10261 gaaaaaaaaa atgagttttg tgaaatagaa ttaaaagaaa atatagtagt tagcttaaat
10321 tgtccaactg gtaaaattaa tccaaaaaat tgttttagaa atgtatatat aaaaagtaat
10381 atgaatgaac aaacaaccga aaatatagaa aatatattta acgaaataaa agttatagat
10441 gcagattatt ttataaataa ttcatcaacc tttttgatga tttccaaaat tacaaaaaaa
10501 gagtttgatt tttattgtac atgtgaagat tataaaacca aaaatatagg aacaatatat
10561 attaaaaatt atgaatatct agattcaaaa cctaaatata aaaataaaca aatttcctat
10621 atagatgtag ttccataccc gcggggaaag ggcg
Restriction sites to linearize plasmid KspI (SacII)
Selectable marker used to select the mutant parasitetgdhfr
Promoter of the selectable markereef1a
Selection (positive) procedurepyrimethamine
Selection (negative) procedureNo
Additional remarks genetic modification
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 PBANKA_0306000
Gene product6-cysteine protein
Gene product: Alternative name230p
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 15'-cccaagcttccgcgggtatatggtaaagaacctactaacac
Additional information primer 1primer L1345: 0.7kb 5'region of PB000214.00.0 (230p gene)
Sequence Primer 25'-cccaagcttgatgtgttttatttggatgtgc
Additional information primer 2primer L1346: 0.7kb 5'region of PB000214.00.0 (230p gene)
Sequence Primer 35'-ccggaattctcttgagcccgttaatg
Additional information primer 3primer L1347: 1kb 3'region of PB000214.00.0 (230p gene)
Sequence Primer 45'-tccccgcgggtatggaactacatctatatagg
Additional information primer 4primer L1348: 1kb 3'region of PB000214.00.0 (230p gene)