Summary

RMgm-8
Malaria parasiteP. berghei
Genotype
Genetic modification not successful
DisruptedGene model (rodent): PBANKA_0417700; Gene model (P.falciparum): PF3D7_0903700; Gene product: alpha tubulin 1
PhenotypeNo phenotype has been described
Last modified: 12 February 2009, 00:28
  *RMgm-8
Successful modificationThe gene/parasite could not be changed/generated by the genetic modification.
The following genetic modifications were attempted Gene disruption
Number of attempts to introduce the genetic modification 2
Reference (PubMed-PMID number) Reference 1 (PMID number) : 16115694
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).
Attempts to generate the mutant parasite were performed by
Name PI/ResearcherT.W. Kooij, C.J. Janse, A.P. Waters
Name Group/DepartmentLeiden Malaria Research Group
Name InstituteLeiden University Medical Center
CityLeiden
CountryThe Netherlands

  Disrupted: Mutant parasite with a disrupted gene
Details of the target gene
Gene Model of Rodent Parasite PBANKA_0417700
Gene Model P. falciparum ortholog PF3D7_0903700
Gene productalpha tubulin 1
Gene product: Alternative name
Details of the genetic modification
Inducable system usedNo
Additional remarks inducable system
Type of plasmid/construct usedPlasmid 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 gtcaacaata aataataaat aaatattgtg gaaataaaat aacatataat
51 tatttttaat acattgattt cccttttatt tttttaaatt tcattgatat
101 aaaaatatat aataataaca tatatgattt caaattaatc ttttcaaaaa
151 tggtgtttat ttttgtatgt ttgtgtatga attaatcaca taacacatct
201 attaaattga gttggtaata tagacacaaa taaatatata tatttttata
251 gcttaaaagt gtgttatgaa tattttaagc atattttctt tttctttgga
301 ttgtgtaaaa tgaactcata taatgcgttt ttttgttttt gttattttgt
351 cattttgtta ttttgctatt ttatggatta atttttgttt ataaaatggg
401 aaataatttt aacatattta aataaatgga gaaaaaatat aaaataatta
451 taaaaaaaag ttaatacaca ttttttcctg ttatagacct tatatttatt
501 tatccatata tatatatata tatatatata tatatatata cataccaagt
551 gaattaagag gaaagctaat ttattattca gaataatata tgaactatat
601 ataattttta ttattttggt gtatattaat ctgtctatat gcatacatgc
651 aataatttat cgacttatat atcaaataac ataaaataga agtgttttaa
701 attatggata tatgctcaat attcattttt tttaataagt tagctatatt
751 taaattatac attttatata tggtctcttt tttttttaaa tattatttaa
801 gtgatcatga aaatataaat aatttttttt tatttaatat ccttttgctt
851 gcatgtggta aatggaaatt tggatgtgtt ttgaargttc ggatatagtt
901 gtatggacat ataatatatt ttgtgaaaaa ttggttttat gtttatactt
951 atgccaatac tttttgagta aaacaaagca agtgcttata aataattaaa
1001 gccaatttta taatatatat ttttttattt aatttgaatt tagtagtata
1051 attttttatg gtaagtgctc aaagagagtt gcttataaag tatggtttgt
1101 ttctttttcg ccattttgaa ttacacatta aaaatatata gatacatata
1151 ttataatatg aaatcattaa taatttaggg aaattctaca aatttaaaaa
1201 cgaataaaat aattgttttt catcatgcca taacacaata ttgatatata
1251 catgtacaaa catttttttt atttggaaaa tataaattat ataaaaaaaa
1301 atgtatagta tacaaaatga gcatattcac acggggtgga cgttcatttt
1351 ttcatttttc ccctgttttt tatgagtata tgataaaatt ttatgaacat
1401 ttacacaaaa tgaaaatgga tatataggaa aaatggagcg gtatttcatt
1451 tatctttgat tgtcatttgg atattatatt accytgggta ggcaattaaa
1501 aatgttaaat aacaatttaa ggaaattata ttttatatat taaaattaac
1551 actgtattat atgattcgct tataaaagcc actctttccc catgcaaagc
1601 tgtttaatat caattttaac aaattacaca catgttaata tatttatata
1651 tataatttat atatttataa tttatatatt tatattttta ttatttatat
1701 atttattatt tattgtgtgt gtcaattcgg gtaggatata cctctttttt
1751 attgtttaaa gcgatttgta ttctaaaata taaagrattt gaaaaagaga
1801 aagatagaat atgatcccat catatatagc cctataattt ttatttagca
1851 gcgaattaat ttttctatta agtttatgtg taattaaaat aacggaatat
1901 atataataca ataaaaaagt gcataaatta aaattttttc aattaaattt
1951 ttttttttaa ggggttatat aatattaaat atataaaata cgattatata
2001 tttttgctac aattttttat attaagatat aaatagtaaa taaatggtat
2051 tatatggcat gtaatatata aattttttcc aatttttatt ttatatacac
2101 ttttcctttt tttgtcataa aacttaaaca atttacacat tcattttaaa
2151 aattgactat ttgtttcaac attttttgag tttccgtttt ataatagtat
2201 tttcatttgt atattgctta tatatataaa tacacaccta aatgttacaa
2251 aggatcaatg cataaaccgg tgtgtctggt cgtcgcgatg acccccaaga
2301 ggggcatcgg catcaacaac ggcctcccgt ggccccactt gaccacagat
2351 ttcaaacact tttctcgtgt gacaaaaacg acgcccgaag aagccagtcg
2401 cctgaacggg tggcttccca ggaaatttgc aaagacgggc gactctggac
2451 ttccctctcc atcagtcggc aagagattca acgccgttgt catgggacgg
2501 aaaacctggg aaagcatgcc tcgaaagttt agacccctcg tggacagatt
2551 gaacatcgtc gtttcctctt ccctcaaaga agaagacatt gcggcggaga
2601 agcctcaagc tgaaggccag cagcgcgtcc gagtctgtgc ttcactccca
2651 gcagctctca gccttctgga ggaagagtac aaggattctg tcgaccagat
2701 ttttgtcgtg ggaggagcgg gactgtacga ggcagcgctg tctctgggcg
2751 ttgcctctca cctgtacatc acgcgtgtag cccgcgagtt tccgtgcgac
2801 gttttcttcc ctgcgttccc cggagatgac attctttcaa acaaatcaac
2851 tgctgcgcag gctgcagctc ctgccgagtc tgtgttcgtt cccttttgtc
2901 cggagctcgg aagagagaag gacaatgaag cgacgtatcg acccatcttc
2951 atttccaaga ccttctcaga caacggggtt ccctacgact ttgtggttct
3001 cgagaagaga aggaagactg acgacgcagc cactgcggaa ccgagcaacg
3051 caatgagctc cttgacgtcc acgagggaga caactcccgt gcacgggttg
3101 caggctcctt cttcggccgc agccattgcc ccggtgttgg cgtggatgga
3151 cgaagaagac cggaaaaaac gcgagcaaaa ggaactgatt cgggccgttc
3201 cgcatgttca ctttagaggc catgaagagt tccagtacct tgatctcatt
3251 gccgacatta ttaacaatgg aaggacaatg gatgaccgaa cgggcgttgg
3301 tgtcatctcc aaattcggct gcactatgcg ctactcgctg gatcaggcct
3351 ttccacttct caccacaaag cgtgtgttct ggaaaggggt cctcgaagag
3401 ttgctgtggt tcattcgcgg cgacacgaac gcaaaccatc tttctgagaa
3451 gggcgtgaag atctgggaca agaatgtgac acgcgagttc ctcgattcgc
3501 gcaatctccc ccaccgagag gtcggagaca tcggcccggg ctacggcttc
3551 cagtggagac acttcggcgc ggcatacaaa gacatgcaca cagactacac
3601 agggcagggc gtcgaccagc tgaagaatgt gatccagatg ctgagaacga
3651 atccaacaga tcgtcgcatg ctcatgactg cctggaatcc tgcagcgctg
3701 gacgaaatgg cgctgccgcc ttgtcacttg ttgtgccagt tctacgtgaa
3751 cgaccagaag gagctgtcgt gcatcatgta tcagcggtcg tgcgatgtcg
3801 gcctcggcgt ccccttcaac atcgcttcct attcgctttt gacgctcatg
3851 gttgcacacg tctgcaacct aaaacctaag gagttcattc acttcatggg
3901 gaacacgcat gtctacacga accatgtcga ggctttaaaa gagcagctgc
3951 ggagagaacc gagaccgttc cccattgtga acatcctcaa caaggaacgc
4001 atcaaggaaa tcgacgattt caccgccgag gattttgagg tcgtgggcta
4051 cgtcccgcac ggacgaatcc agatggagat ggctgtctag cggaaataca
4101 gaagctggct ttgatcccgt ttttcttact tatatattta taccaattga
4151 ttgtatttat aactgtaaaa atgtgtatgt tgtgtgcata tttttttttg
4201 tgcatgcaca tgcatgtaaa tagctaaaat tatgaacatt ttattttttg
4251 ttcagaaaaa aaaaacttta cacacataaa atggctagta tgaatagcca
4301 tattttatat aaattaaatc ctatgaattt atgaccatat taaaaattta
4351 gatatttatg gaacataata tgtttgaaac aataagacaa aattattatt
4401 attattatta tttttactgt tataattatg ttgtctcttc aatgattcat
4451 aaatagttgg acttgatttt taaaatgttt ataatatgat tagcatagtt
4501 aaataaaaaa agttgaaaaa ttaaaaaaaa acatataaac acaaatgatg
4551 ttttttcctt caatttcgat atccatcacc acaagtttca actgctgtag
4601 ttgaaccata taattcagtt ttatcaacac attcattatt agaacacaca
4651 gatgtagcta ttatgctaga taatgaagct atttacgata tatgcaaaaa
4701 aaatttagat atagaaagac ctacttatac aaatttaaat agattgattg
4751 cacaagttat atcatcttta acagcttcat tacgttttga tggtgcatta
4801 aatgttgatg ttaccgaatt tcaaactaat ttagtaccat atccacgtat
4851 tcattttatg ttatcatcat atgcccctgt agttagtgct gaaaaagcat
4901 atcatgaaca attgtctgta tcagaaatta caaactcagc atttgaacca
4951 gcaaatatga tggcaaaatg tgatcctaga catggaaaat atatggcttg
5001 ttgtttaatg tatagaggag atgtagtacc aaaagatgtg aatgccgctg
5051 ttgccactat aaaaacgaaa agaactattc aatttgttga ttggtgccca
5101 actggattta aatgtggtat taattatcaa cctccaacag tagtacccgg
5151 tggagactta gcaaaagtta tgagagctgt ttgtatgatt tctaactcaa
5201 ctgctattgc tgaagtttga attcctgcag cccgggggat ccactagttc
5251 tagagcggcc gccaccgcgg tggagctcca gcttttgttc cctttagtga
5301 gggttaattt cgagcttggc gtaatcatgg tcatagctgt ttcctgtgtg
5351 aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa
5401 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg
5451 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
5501 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct
5551 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
5601 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
5651 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
5701 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
5751 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
5801 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
5851 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
5901 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
5951 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
6001 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
6051 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
6101 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
6151 ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg
6201 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
6251 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
6301 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
6351 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
6401 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
6451 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
6501 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
6551 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
6601 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
6651 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
6701 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
6751 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
6801 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
6851 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
6901 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
6951 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
7001 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
7051 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
7101 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
7151 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
7201 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
7251 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
7301 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
7351 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
7401 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
7451 tccgcgcaca tttccccgaa aagtgccacc taaattgtaa gcgttaatat
7501 tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc
7551 aataggccga aatcggcaaa atcccttata aatcaaaaga atagaccgag
7601 atagggttga gtgttgttcc agtttggaac aagagtccac tattaaagaa
7651 cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc
7701 cactacgtga accatcaccc taatcaagtt ttttggggtc gaggtgccgt
7751 aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg
7801 gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag
7851 cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc
7901 acacccgccg cgcttaatgc gccgctacag ggcgcgtccc attcgccatt
7951 caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
8001 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta
8051 acgccagggt tttcccagtc acgacgttgt aaaacgacgg ccagtgaatt
8101 gtaatacgac tcactatagg gcgaattggg taccgggccc cccctcgagg
8151 tcgacggtat cgataagctt ggatcccaca gcatatgcta attatatata
8201 cttataatat acagaattta tatatatcgt gattgcttgt ctgaataccc
8251 ttaacaaata aaaattctta taagtgcacc acttagctta catcctgaca
8301 gaattttttt tatatttata agaaaaaatt ataatataat ttacaccaaa
8351 tatgcgtgta aaaaccaaca atatatgcat agtgtattta atccatttaa
8401 aggaaaacac aatatatatt tattttcacc gcgataaaag tataagaaaa
8451 aaaaatattg cataaaatac tagcattaca tatttgtgaa tttttttttc
8501 tttttttact tcaaaaaata ttgttatttt gattttacat acattattta
8551 aataaatgaa attgagagta ttatattatt ttatatatat gtgtgagctc
8601 tcaacctcaa tgttttaagt atgcatagca attttttaca agtaattaat
8651 aaattttacc cgaattaaaa aatattacaa aatacaaaaa ttagaacaaa
8701 atataagtaa atatatatac acatataaac aaatatatat atgtataata
8751 tcattaatta acatattatc acatatttta tttactgttt tatatttaat
8801 ttttactata ttttaaaaac aattgtttat tttataatat acaagtaaaa
8851 atgagagaag taataagtat acatgaagct tgcatgcctg cag
"
Restriction sites to linearize plasmid
Partial or complete disruption of the geneComplete
Additional remarks partial/complete disruption
Selectable marker used to select the mutant parasitetgdhfr
Promoter of the selectable markerpbdhfr
Selection (positive) procedurepyrimethamine
Selection (negative) procedureNo
Additional remarks genetic modificationAlpha-tubulin I is one of the two alpha-tubulin genes (I and II) in Plasmodium. This gene is highly expressed in all blood stages (higher expression in females compared to males). Expression was also observed in ookinetes, oocysts and sporozoites (additional P. berghei gene models: PB300531.00.0)
Additional remarks selection procedure
Primer information: Primers used for amplification of the target sequences  Click to view information
Primer information: Primers used for amplification of the target sequences  Click to hide information
Sequence Primer 1CCCAAGCTTGGATCCCACAGCATATGCTAATTATATAT
Additional information primer 1Primer L466; 5'UTR Pb alpha-tub I
Sequence Primer 2CCCAAGCTTCATGTATACTTATTACTTCTCTC
Additional information primer 2Primer L467; 5'UTR Pb alpha-tub I
Sequence Primer 3GGATATCCATCACCACAGGTTTCTACTGC
Additional information primer 3Primer L470; Exon 3 Pb alpha-tub I & II
Sequence Primer 4GGAATTCAAACTTCAGCAATAGCAGTTGAG
Additional information primer 4Primer L471; Exon 3 Pb alpha-tub I & II
Sequence Primer 5
Additional information primer 5
Sequence Primer 6
Additional information primer 6