Summary

RMgm-350
Malaria parasiteP. berghei
Genotype
DisruptedGene model (rodent): PBANKA_0306100; Gene model (P.falciparum): PF3D7_0209000; Gene product: 6-cysteine protein | transmission-blocking target antigen s230 (P230)
Phenotype Gametocyte/Gamete; Fertilization and ookinete;
Last modified: 15 April 2010, 07:16
  *RMgm-350
Successful modificationThe parasite was generated by the genetic modification
The mutant contains the following genetic modification(s) Gene disruption
Reference (PubMed-PMID number) Reference 1 (PMID number) : 20386715
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/ResearcherM.R. van Dijk; C.J. Janse
Name Group/DepartmentLeiden Malaria Research Group
Name InstituteLeiden University Medical Center
CityLeiden
CountryThe Netherlands
Name of the mutant parasite
RMgm numberRMgm-350
Principal name310cl1; 323cl1
Alternative nameΔp230
Standardized name
Is the mutant parasite cloned after genetic modificationYes
Phenotype
Asexual blood stageNot different from wild type
Gametocyte/GameteNormal numbers of male and female gametocytes are produced. Male and female gamete formation is normal (escape from host red blood cell, formation of 8 motile male gametes). Male gametes are strongly affected in their fertility, resulting in nearly complete inhibition of ookinete formation in vitro (>0.99%). Motile males fail to attach to and penetrate female gametes.
Mutant female gametes are fertile as shown by cross-fertilisation with wild type male gametes.
Fertilization and ookineteMale gametes are strongly affected in their fertility, resulting in nearly complete inhibition of ookinete formation in vitro (>0.99%); Motile males fail to attach to and penetrate female gametes.
Mutant female gametes are fertile as shown by cross-fertilization with wild type male gametes.
Mutant parasites produce low numbers of ookinetes in vivo (Anopheles stephensi). See also 'Additional remarks phenotype'.
OocystNot different from wild type
SporozoiteNot different from wild type
Liver stageNot different from wild type
Additional remarks phenotype

Mutant/mutation
The mutant lacks expression of P230.

Protein (function)
The P230 protein is a member of a small family of proteins, the 6-cysteine (cys) family of (surface) proteins. The proteins are characterized by domains of roughly 120 amino acids in size that contain six positionally conserved cysteines (6-cys). Although some species of Plasmodium (may) contain unique members of the 6-cys family, ten members have been identified that are conserved both in structure as well as in genome organization throughout the genus. Some of the conserved 6-cys proteins are encoded by genes that form paralogous gene-pairs which are closely linked in the genome separated by less then 2 kb of intergenic region. Most members have a GPI anchor and are predicted membrane surface proteins whereas others appear to be secreted and most members are expressed in a discrete stage-specific manner in gametocytes, sporozoites or merozoites (see also 'Additional Information'). Evidence has been presented that P230 is specifically expressed in male gametocytes/gametes of P. berghei. P230 does not have a GPI anchor. In P. falciparum P230 has been shown to be located on the surface of male and female gametes and evidence has been found that P230 forms a complex with P48/45 at the surface of gametocytes and gametes.

Phenotype
Phenotype analyses demonstrate that P230 plays an important role in fertilization (male fertility factor). Motile males fail to attach to and penetrate female gametes (see phenotype description). Mutant female gametes are fertile as has been shown by cross-fertilisation with wild type male gametes. This phenotype is comparable to the phenotype of mutants lacking expression of P48/45 (see RMgm-15).

Additional information
The mutant  shows very low levels of fertilization with higher fertilization rates in the mosquito than in in vitro ookinete cultures. Mutant parasites produce low numbers of ookinetes and oocysts in Anopheles stephensi mosquitoes (see also the phenotype description 'Fertilization and ookinete'). Both the ookinetes and oocysts show a normal morphology and oocysts produce normal numbers of sporozoites that are infectious to mice.

The p230 gene forms a paralogous gene pair with the gene p230p (PB000403.00.0; PFB0400w), which are closely linked in the genome.

In P. falciparum it has been shown that male gametes with a disrupted p230 gene are incapable of interacting with erythrocytes and do not form the characteristic exflagellation centers and these mutants show a strong reduction in oocyst formation (Eksi et al, 2006, Mol. Microbiol. 61, 991-998). In the P. berghei mutants lacking P230, analyses of both exflagellating microgametocytes and free, motile male gametes in live preparations did not reveal any difference in their capacity to interact with red blood cells compared to wild type gametes and normal exflagellation centers were observed.

Table: P. falciparum gene members of the 6-cys family

Gene P. falciparum Gene P. falciparum
p48/45  PF13_0247 p12p  PFF0620c
p47  PF13_0248 p230p  PFB0400w
p36  PFD0210c p230  PFB0405w
p52  PFD0215c p38  PFE0395c
p12  PFF0615c p41  PFD0240c


Other mutants


  Disrupted: Mutant parasite with a disrupted gene
Details of the target gene
Gene Model of Rodent Parasite PBANKA_0306100
Gene Model P. falciparum ortholog PF3D7_0209000
Gene product6-cysteine protein | transmission-blocking target antigen s230
Gene product: Alternative nameP230
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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GGGCCCCCCCTCGAGGTCGACGGTATCGATAAGCTTCATATTTTCCTAAAAGAGCTCCAT
ATATTGTTTTAGAACTAACTAAAAAAACAGAAAAAGAAAGCCAAACAAAATTGAAAGAAA
AAAAAATAGCTGATTTATTATATGGTACTATCATACCTAAAATACATAATGGAAAAAATA
ATAATTTTAATCGCGGATCTATAAGATTTATATTGCCTCCAATTGTTGTTAAGCCTACAA
CCTTTTATATAATATGTGATAATTCAAGATCCAAAAAAAATTCATTTAGAGGAAAAAAAG
GAATTGTAAAAATACATGTTGCACCTTATGGCAATATTGAAAAAGGATGTAACTACAGTA
AAAACAAAAATCACTTATTTGGAATAAATATAAATAATTATGAGAAAGAAAAACAAATAG
ATGAAATTGAAAATAGAAAAGGAAACCTTCCATGTTTTTTTCAATTAAATGCAGGGGAAG
CTGGAGGTATTATTTTTCCAGAAAACTATAAATCTAGTACATGTTTTGAAGAAGTAGTAG
AATATAATCCTAATAAAAACTGGAATAGAAAAAAAAAAGAAATAAATGAACTAATAAATG
GTGCTATAGTATATAAAAGTGATATAAAATATAAATATTTTAATGCAATATACATTGGGA
TACCTACTAGCTATAGAAAATCATTACATATTTTTTGCAACATTACAACAACAGATAATA
GTTTTCATATGGTTTATCTAAGTATCAATCAAGAGATAAATATATCTGTTTTTGATTTAT
TTGAATCTTTTATACGATTAAATAAAATGACAGAAGTAGCTCAAAAAGTTGATCAAAAAA
ATGAATACATATGTGATTTTACAAAAAGATTAGATGCACCGCATGATAATAATGAAAATA
ATAAAAAAGTTATTATATGTAGAAAAAAACTAAAAGAATTTGATATATTTAGATTAAAAT
GTGATTTTAATAAAACAAAAAATAAAAATATTGGCATGATTCCAGGTTCGTTAAAGGAAG
CTTGCATGCCTGCAGGTCAACAATAAATAATAAATAAATATTGTGGAAATAAAATAACAT
ATAATTATTTTTAATACATTGATTTCCCTTTTATTTTTTTAAATTTCATTGATATAAAAA
TATATAATAATAACATATATGATTTCAAATTAATCTTTTCAAAAATGGTGTTTATTTTTG
TATGTTTGTGTATGAATTAATCACATAACACATCTATTAAATTGAGTTGGTAATATAGAC
ACAAATAAATATATATATTTTTATAGCTTAAAAGTGTGTTATGAATATTTTAAGCATATT
TTCTTTTTCTTTGGATTGTGTAAAATGAACTCATATAATGCGTTTTTTTGTTTTTGTTAT
TTTGTCATTTTGTTATTTTGCTATTTTATGGATTAATTTTTGTTTATAAAATGGGAAATA
ATTTTAACATATTTAAATAAATGGAGAAAAAATATAAAATAATTATAAAAAAAAGTTAAT
ACACATTTTTTCCTGTTATAGACCTTATATTTATTTATCCATATATATATATATATATAT
ATATATATATATATACATACCAAGTGAATTAAGAGGAAAGCTAATTTATTATTCAGAATA
ATATATGAACTATATATAATTTTTATTATTTTGGTGTATATTAATCTGTCTATATGCATA
CATGCAATAATTTATCGACTTATATATCAAATAACATAAAATAGAAGTGTTTTAAATTAT
GGATATATGCTCAATATTCATTTTTTTTAATAAGTTAGCTATATTTAAATTATACATTTT
ATATATGGTCTCTTTTTTTTTTAAATATTATTTAAGTGATCATGAAAATATAAATAATTT
TTTTTTATTTAATATCCTTTTGCTTGCATGTGGTAAATGGAAATTTGGATGTGTTTTGAA
RGTTCGGATATAGTTGTATGGACATATAATATATTTTGTGAAAAATTGGTTTTATGTTTA
TACTTATGCCAATACTTTTTGAGTAAAACAAAGCAAGTGCTTATAAATAATTAAAGCCAA
TTTTATAATATATATTTTTTTATTTAATTTGAATTTAGTAGTATAATTTTTTATGGTAAG
TGCTCAAAGAGAGTTGCTTATAAAGTATGGTTTGTTTCTTTTTCGCCATTTTGAATTACA
CATTAAAAATATATAGATACATATATTATAATATGAAATCATTAATAATTTAGGGAAATT
CTACAAATTTAAAAACGAATAAAATAATTGTTTTTCATCATGCCATAACACAATATTGAT
ATATACATGTACAAACATTTTTTTTATTTGGAAAATATAAATTATATAAAAAAAAATGTA
TAGTATACAAAATGAGCATATTCACACGGGGTGGACGTTCATTTTTTCATTTTTCCCCTG
TTTTTTATGAGTATATGATAAAATTTTATGAACATTTACACAAAATGAAAATGGATATAT
AGGAAAAATGGAGCGGTATTTCATTTATCTTTGATTGTCATTTGGATATTATATTACCYT
GGGTAGGCAATTAAAAATGTTAAATAACAATTTAAGGAAATTATATTTTATATATTAAAA
TTAACACTGTATTATATGATTCGCTTATAAAAGCCACTCTTTCCCCATGCAAAGCTGTTT
AATATCAATTTTAACAAATTACACACATGTTAATATATTTATATATATAATTTATATATT
TATAATTTATATATTTATATTTTTATTATTTATATATTTATTATTTATTGTGTGTGTCAA
TTCGGGTAGGATATACCTCTTTTTTATTGTTTAAAGCGATTTGTATTCTAAAATATAAAG
RATTTGAAAAAGAGAAAGATAGAATATGATCCCATCATATATAGCCCTATAATTTTTATT
TAGCAGCGAATTAATTTTTCTATTAAGTTTATGTGTAATTAAAATAACGGAATATATATA
ATACAATAAAAAAGTGCATAAATTAAAATTTTTTCAATTAAATTTTTTTTTTTAAGGGGT
TATATAATATTAAATATATAAAATACGATTATATATTTTTGCTACAATTTTTTATATTAA
GATATAAATAGTAAATAAATGGTATTATATGGCATGTAATATATAAATTTTTTCCAATTT
TTATTTTATATACACTTTTCCTTTTTTTGTCATAAAACTTAAACAATTTACACATTCATT
TTAAAAATTGACTATTTGTTTCAACATTTTTTGAGTTTCCGTTTTATAATAGTATTTTCA
TTTGTATATTGCTTATATATATAAATACACACCTAAATGTTACAAAGGATCAATGCATAA
ACCGGTGTGTCTGGTCGTCGCGATGACCCCCAAGAGGGGCATCGGCATCAACAACGGCCT
CCCGTGGCCCCACTTGACCACAGATTTCAAACACTTTTCTCGTGTGACAAAAACGACGCC
CGAAGAAGCCAGTCGCCTGAACGGGTGGCTTCCCAGGAAATTTGCAAAGACGGGCGACTC
TGGACTTCCCTCTCCATCAGTCGGCAAGAGATTCAACGCCGTTGTCATGGGACGGAAAAC
CTGGGAAAGCATGCCTCGAAAGTTTAGACCCCTCGTGGACAGATTGAACATCGTCGTTTC
CTCTTCCCTCAAAGAAGAAGACATTGCGGCGGAGAAGCCTCAAGCTGAAGGCCAGCAGCG
CGTCCGAGTCTGTGCTTCACTCCCAGCAGCTCTCAGCCTTCTGGAGGAAGAGTACAAGGA
TTCTGTCGACCAGATTTTTGTCGTGGGAGGAGCGGGACTGTACGAGGCAGCGCTGTCTCT
GGGCGTTGCCTCTCACCTGTACATCACGCGTGTAGCCCGCGAGTTTCCGTGCGACGTTTT
CTTCCCTGCGTTCCCCGGAGATGACATTCTTTCAAACAAATCAACTGCTGCGCAGGCTGC
AGCTCCTGCCGAGTCTGTGTTCGTTCCCTTTTGTCCGGAGCTCGGAAGAGAGAAGGACAA
TGAAGCGACGTATCGACCCATCTTCATTTCCAAGACCTTCTCAGACAACGGGGTTCCCTA
CGACTTTGTGGTTCTCGAGAAGAGAAGGAAGACTGACGACGCAGCCACTGCGGAACCGAG
CAACGCAATGAGCTCCTTGACGTCCACGAGGGAGACAACTCCCGTGCACGGGTTGCAGGC
TCCTTCTTCGGCCGCAGCCATTGCCCCGGTGTTGGCGTGATGGACGAAGAAGACCGGAAA
AAACGCGAGCAAAAGGAACTGATTCGGGCCGTTCCGCATGTTCACTTTAGAGGCCATGAA
GAGTTCCAGTACCTTGATCTCATTGCCGACATTATTAACAATGGAAGGACAATGGATGAC
CGAACGGGCGTTGGTGTCATCTCCAAATTCGGCTGCACTATGCGCTACTCGCTGGATCAG
GCCTTTCCACTTCTCACCACAAAGCGTGTGTTCTGGAAAGGGGTCCTCGAAGAGTTGCTG
TGGTTCATTCGCGGCGACACGAACGCAAACCATCTTTCTGAGAAGGGCGTGAAGATCTGG
GACAAGAATGTGACACGCGAGTTCCTCGATTCGCGCAATCTCCCCCACCGAGAGGTCGGA
GACATCGGCCCGGGCTACGGCTTCCAGTGGAGACACTTCGGCGCGGCATACAAAGACATG
CACACAGACTACACAGGGCAGGGCGTCGACCAGCTGAAGAATGTGATCCAGATGCTGAGA
ACGAATCCAACAGATCGTCGCATGCTCATGACTGCCTGGAATCCTGCAGCGCTGGACGAA
ATGGCGCTGCCGCCTTGTCACTTGTTGTGCCAGTTCTACGTGAACGACCAGAAGGAGCTG
TCGTGCATCATGTATCAGCGGTCGTGCGATGTCGGCCTCGGCGTCCCCTTCAACATCGCT
TCCTATTCGCTTTTGACGCTCATGGTTGCACACGTCTGCAACCTAAAACCTAAGGAGTTC
ATTCACTTCATGGGGAACACGCATGTCTACACGAACCATGTCGAGGCTTTAAAAGAGCAG
CTGCGGAGAGAACCGAGACCGTTCCCCATTGTGAACATCCTCAACAAGGAACGCATCAAG
GAAATCGACGATTTCACCGCCGAGGATTTTGAGGTCGTGGGCTACGTCCCGCACGGACGA
ATCCAGATGGAGATGGCTGTCTAGCGGAAATACAGAAGCTAGCTTTGATCCCGTTTTTCT
TACTTATATATTTATACCAATTGATTGTATTTATAACTGTAAAAATGTGTATGTTGTGTG
CATATTTTTTTTTGTGCATGCACATGCATGTAAATAGCTAAAATTATGAACATTTTATTT
TTTGTTCAGAAAAAAAAAACTTTACACACATAAAATGGCTAGTATGAATAGCCATATTTT
ATATAAATTAAATCCTATGAATTTATGACCATATTAAAAATTTAGATATTTATGGAACAT
AATATGTTTGAAACAATAAGACAAAATTATTATTATTATTATTATTTTTACTGTTATAAT
TATGTTGTCTCTTCAATGATTCATAAATAGTTGGACTTGATTTTTAAAATGTTTATAATA
TGATTAGCATAGTTAAATAAAAAAAGTTGAAAAATTAAAAAAAAACATATAAACACAAAT
GATGTTTTTTCCTTCAATTTCGATATCGGAAAATATTTTAATGAATCTCCTAATAAAAAT
GGTTTATTAGTAAATTTATTAACATACTTAAATTCATTAGATGTATTATCTATGAATACA
GAAATAGAGTCTATAAATATAGATGTAAATGATTCTGATCAAAATATTGTAGATATATCA
TTTCAAGTACCAGCCTATATATATAATACCAATCCATACTATTTTGTGTTTGGATGTAAT
AATACCAGAGATAATGGGAAAATCGGTATTGTTGAATTAATTATTTCAAAAAATGAAGAA
ATGATAAAAGGTTGTAATTTTAATTCTGATGCTATAGAACATTTTTCAAACAACATGAGA
CCGGATGAGACCGAATGTAAAATAGATGCATGTCCAAATGATATTATTGGTTTTATTTGC
CCCAAAAAACAGAATTTTGTTTCATCCAAGCACGTCCTAGATATAGATGCAGATACAGAT
GCAGATTTAGAGAATGTGGACGTCAACCCAAATGACTGTTTTGATTCTATAAATATAGAT
TCTACTAAAAAATATATAGTGAATGAATTGCCAGGAGCCCAAACATATCGTAACAAGTCA
AGAAATATGCCAAGGTATTTTAAAGTGCCATATCATAACAATGAATTAGATGTTATATTT
CAGTGCTCATGTGTAATGGGTTCTAAAACCAATAAAATAATAGTAACCGTGAAAGCCTTA
AATGGCCAAATCCCGAAAAAATATGAAAAAAGTGAAATAAAAAGTAGCCCATCAATAGTA
GATGGCCCTGTACGTGCGTGTCCTCATAAACAATCTATAGAACCAAAGTTAATAAAATGG
AATAATAAAGATACTGAAACTATAGATAGTTGTAAAATAGATGTAAACAGATTCTATTCG
TATGCTCAATTAGCATGCCCAAACAATAATTTTTCTATGCATAGCAATATTCAGTTAACT
TACAACATATTAAAACCCAATAAAATAGATGAATATAAAACTTTTAATGAATCTGAATTA
AATAAGCTAATTCCACATTCAGAAATACGGATCCACTAGTTCTAGAGCGGCCGCCACCGC
GGTGGAGCTCCAATTCGCCCTATAGTGAGTCGTATTACGCGCGCTCACTGGCCGTCGTTT
TACAACGTCGTGACTGGGAAAACCCTGGCGTTACCCAACTTAATCGCCTTGCAGCACATC
CCCCTTTCGCCAGCTGGCGTAATAGCGAAGAGGCCCGCACCGATCGCCCTTCCCAACAGT
TGCGCAGCCTGAATGGCGAATGGGACGCGCCCTGTAGCGGCGCATTAAGCGCGGCGGGTG
TGGTGGTTACGCGCAGCGTGACCGCTACACTTGCCAGCGCCCTAGCGCCCGCTCCTTTCG
CTTTCTTCCCTTCCTTTCTCGCCACGTTCGCCGGCTTTCCCCGTCAAGCTCTAAATCGGG
GGCTCCCTTTAGGGTTCCGATTTAGTGCTTTACGGCACCTCGACCCCAAAAAACTTGATT
AGGGTGATGGTTCACGTAGTGGGCCATCGCCCTGATAGACGGTTTTTCGCCCTTTGACGT
TGGAGTCCACGTTCTTTAATAGTGGACTCTTGTTCCAAACTGGAACAACACTCAACCCTA
TCTCGGTCTATTCTTTTGATTTATAAGGGATTTTGCCGATTTCGGCCTATTGGTTAAAAA
ATGAGCTGATTTAACAAAAATTTAACGCGAATTTTAACAAAATATTAACGCTTACAATTT
AGGTGGCACTTTTCGGGGAAATGTGCGCGGAACCCCTATTTGTTTATTTTTCTAAATACA
TTCAAATATGTATCCGCTCATGAGACAATAACCCTGATAAATGCTTCAATAATATTGAAA
AAGGAAGAGTATGAGTATTCAACATTTCCGTGTCGCCCTTATTCCCTTTTTTGCGGCATT
TTGCCTTCCTGTTTTTGCTCACCCAGAAACGCTGGTGAAAGTAAAAGATGCTGAAGATCA
GTTGGGTGCACGAGTGGGTTACATCGAACTGGATCTCAACAGCGGTAAGATCCTTGAGAG
TTTTCGCCCCGAAGAACGTTTTCCAATGATGAGCACTTTTAAAGTTCTGCTATGTGGCGC
GGTATTATCCCGTATTGACGCCGGGCAAGAGCAACTCGGTCGCCGCATACACTATTCTCA
GAATGACTTGGTTGAGTACTCACCAGTCACAGAAAAGCATCTTACGGATGGCATGACAGT
AAGAGAATTATGCAGTGCTGCCATAACCATGAGTGATAACACTGCGGCCAACTTACTTCT
GACAACGATCGGAGGACCGAAGGAGCTAACCGCTTTTTTGCACAACATGGGGGATCATGT
AACTCGCCTTGATCGTTGGGAACCGGAGCTGAATGAAGCCATACCAAACGACGAGCGTGA
CACCACGATGCCTGTAGCAATGGCAACAACGTTGCGCAAACTATTAACTGGCGAACTACT
TACTCTAGCTTCCCGGCAACAATTAATAGACTGGATGGAGGCGGATAAAGTTGCAGGACC
ACTTCTGCGCTCGGCCCTTCCGGCTGGCTGGTTTATTGCTGATAAATCTGGAGCCGGTGA
GCGTGGGTCTCGCGGTATCATTGCAGCACTGGGGCCAGATGGTAAGCCCTCCCGTATCGT
AGTTATCTACACGACGGGGAGTCAGGCAACTATGGATGAACGAAATAGACAGATCGCTGA
GATAGGTGCCTCACTGATTAAGCATTGGTAACTGTCAGACCAAGTTTACTCATATATACT
TTAGATTGATTTAAAACTTCATTTTTAATTTAAAAGGATCTAGGTGAAGATCCTTTTTGA
TAATCTCATGACCAAAATCCCTTAACGTGAGTTTTCGTTCCACTGAGCGTCAGACCCCGT
AGAAAAGATCAAAGGATCTTCTTGAGATCCTTTTTTTCTGCGCGTAATCTGCTGCTTGCA
AACAAAAAAACCACCGCTACCAGCGGTGGTTTGTTTGCCGGATCAAGAGCTACCAACTCT
TTTTCCGAAGGTAACTGGCTTCAGCAGAGCGCAGATACCAAATACTGTCCTTCTAGTGTA
GCCGTAGTTAGGCCACCACTTCAAGAACTCTGTAGCACCGCCTACATACCTCGCTCTGCT
AATCCTGTTACCAGTGGCTGCTGCCAGTGGCGATAAGTCGTGTCTTACCGGGTTGGACTC
AAGACGATAGTTACCGGATAAGGCGCAGCGGTCGGGCTGAACGGGGGGTTCGTGCACACA
GCCCAGCTTGGAGCGAACGACCTACACCGAACTGAGATACCTACAGCGTGAGCTATGAGA
AAGCGCCACGCTTCCCGAAGGGAGAAAGGCGGACAGGTATCCGGTAAGCGGCAGGGTCGG
AACAGGAGAGCGCACGAGGGAGCTTCCAGGGGGAAACGCCTGGTATCTTTATAGTCCTGT
CGGGTTTCGCCACCTCTGACTTGAGCGTCGATTTTTGTGATGCTCGTCAGGGGGGCGGAG
CCTATGGAAAAACGCCAGCAACGCGGCCTTTTTACGGTTCCTGGCCTTTTGCTGGCCTTT
TGCTCACATGTTCTTTCCTGCGTTATCCCCTGATTCTGTGGATAACCGTATTACCGCCTT
TGAGTGAGCTGATACCGCTCGCCGCAGCCGAACGACCGAGCGCAGCGAGTCAGTGAGCGA
GGAAGCGGAAGAGCGCCCAATACGCAAACCGCCTCTCCCCGCGCGTTGGCCGATTCATTA
ATGCAGCTGGCACGACAGGTTTCCCGACTGGAAAGCGGGCAGTGAGCGCAACGCAATTAA
TGTGAGTTAGCTCACTCATTAGGCACCCCAGGCTTTACACTTTATGCTTCCGGCTCGTAT
GTTGTGTGGAATTGTGAGCGGATAACAATTTCACACAGGAAACAGCTATGACCATGATTA
CGCCAAGCTCGAAATTAACCCTCACTAAAGGGAACAAAAGCTGGTACC
Restriction sites to linearize plasmid ClaI, BamHI
Partial or complete disruption of the genePartial
Additional remarks partial/complete disruption Of the 4486bp coding region, nucleotides 831-1410 were disrupted
Selectable marker used to select the mutant parasitetgdhfr
Promoter of the selectable markerpbdhfr
Selection (positive) procedurepyrimethamine
Selection (negative) procedureNo
Additional remarks genetic modification
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 1L925 (HindIII); Pb230 5'end sense
Additional information primer 1CCCAAGCTTCATATTTTCCTAAAAGAGCTCC
Sequence Primer 2L1360 (HindIII); Pbs230R1 5'KO
Additional information primer 2CCCAAGCTTCACTTTTATATACTATAGCACC
Sequence Primer 3L1357 (EcoRV); Pbs230F 3'KO
Additional information primer 3GGATATCGGAAAATATTTTAATGAATCCCC
Sequence Primer 4L1358 (BamHI); Pbs230R 3'KO
Additional information primer 4CGCGGATCCGTATTTCTGAATGTGGAATTAG
Sequence Primer 5
Additional information primer 5
Sequence Primer 6
Additional information primer 6