Summary

RMgm-15
Malaria parasiteP. berghei
Genotype
DisruptedGene model (rodent): PBANKA_1359600; Gene model (P.falciparum): PF3D7_1346700; Gene product: transmission blocking target antigen precursor|6-cysteine protein (P48/45)
Phenotype Gametocyte/Gamete; Fertilization and ookinete;
Last modified: 24 December 2009, 22:25
  *RMgm-15
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) : 11163248
MR4 number MRA-869
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, A.P. Waters
Name Group/DepartmentLeiden Malaria Research Group
Name InstituteLeiden University Medical Center
CityLeiden
CountryThe Netherlands
Name of the mutant parasite
RMgm numberRMgm-15
Principal name120cl1; 120cl2; 137cl7; 137cl8
Alternative namep48/45-
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 the P48/45 protein.

Protein/function
The P48/45 protein (PF13_0247) is a member of a small family of proteins, the 6-cysteine (cys) family of (surface) proteins (Thompson J. et al., Mol. Biochem. Parasitol. (2001)118, 147-54). 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 (Thompson et al., 2001). 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.
P48/45 is a surface protein of both male and female gametes and is considered as a major candidate antigen for development of a transmission blocking vaccine.

Phenotype(function)
Phenotype analyses demonstrate that P48/45 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.

Additional information
The function of the protein in fertilization is conserved between P. falciparum and P. berghei (van Dijk et al., Cell (2001), 104, 153-164).
This mutant is frequently used in cross-fertilization studies in which the fertile females are used to test the fertility of males of other mutants (Khan et al., Cell (2005) 121, 675-87; Raine, J.D. et al., PLoS Pathog (2007) 3,e30).
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 p48/45 gene forms a paralogous gene pair with the gene p47 that encodes P47 (PB001526.02.0; PF13_0248, pf47), which are closely linked in the genome. 

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


Additional mutants
RMgm-345: The mutant lacks expression of the P48/45 protein and expresses GFP under the control of a male specific promoter and RFP under the control of a female specific promoter. This mutant has been generated to be able to distinguish the female gametocytes/gametes and ookinetes based on their RFP expression in cross-fertilisation experiments with fertile males of other mutants and for determination of meiosis in cross-fertilisation studies. Meiosis in zygotes is determined by FACS analysis of the DNA content of Hoechst-stained gametes/zygotes selected based on their RFP-fluorescence.
RMgm-346: An independent mutant lacking expression of P48/45 which expresses GFP under the constitutive eef1a promoter. This mutant shows the same phenotype as the mutant described here. This mutant has been generated to be able to distinguish the female gametocytes and ookinetes based on their high GFP expression in cross-fertilisation experiments with fertile males of other mutants (see above).
RMgm-349: A mutant lacking expression of both P47 and P48/45
 


  Disrupted: Mutant parasite with a disrupted gene
Details of the target gene
Gene Model of Rodent Parasite PBANKA_1359600
Gene Model P. falciparum ortholog PF3D7_1346700
Gene producttransmission blocking target antigen precursor|6-cysteine protein
Gene product: Alternative nameP48/45
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
Plasmid/construct sequence
Click to view information
Click to hide information
" 1 agcttgcatg cctgcaggtc aacaataaat aataaataaa tattgtggaa
51 ataaaataac atataattat ttttaataca ttgatttccc ttttattttt
101 ttaaatttca ttgatataaa aatatataat aataacatat atgatttcaa
151 attaatcttt tcaaaaatgg tgtttatttt tgtatgtttg tgtatgaatt
201 aatcacataa cacatctatt aaattgagtt ggtaatatag acacaaataa
251 atatatatat ttttatagct taaaagtgtg ttatgaatat tttaagcata
301 ttttcttttt ctttggattg tgtaaaatga actcatataa tgcgtttttt
351 tgtttttgtt attttgtcat tttgttattt tgctatttta tggattaatt
401 tttgtttata aaatgggaaa taattttaac atatttaaat aaatggagaa
451 aaaatataaa ataattataa aaaaaagtta atacacattt tttcctgtta
501 tagaccttat atttatttat ccatatatat atatatatat atatatatat
551 atatatacat accaagtgaa ttaagaggaa agctaattta ttattcagaa
601 taatatatga actatatata atttttatta ttttggtgta tattaatctg
651 tctatatgca tacatgcaat aatttatcga cttatatatc aaataacata
701 aaatagaagt gttttaaatt atggatatat gctcaatatt catttttttt
751 aataagttag ctatatttaa attatacatt ttatatatgg tctctttttt
801 ttttaaatat tatttaagtg atcatgaaaa tataaataat ttttttttat
851 ttaatatcct tttgcttgca tgtggtaaat ggaaatttgg atgtgttttg
901 aargttcgga tatagttgta tggacatata atatattttg tgaaaaattg
951 gttttatgtt tatacttatg ccaatacttt ttgagtaaaa caaagcaagt
1001 gcttataaat aattaaagcc aattttataa tatatatttt tttatttaat
1051 ttgaatttag tagtataatt ttttatggta agtgctcaaa gagagttgct
1101 tataaagtat ggtttgtttc tttttcgcca ttttgaatta cacattaaaa
1151 atatatagat acatatatta taatatgaaa tcattaataa tttagggaaa
1201 ttctacaaat ttaaaaacga ataaaataat tgtttttcat catgccataa
1251 cacaatattg atatatacat gtacaaacat tttttttatt tggaaaatat
1301 aaattatata aaaaaaaatg tatagtatac aaaatgagca tattcacacg
1351 gggtggacgt tcattttttc atttttcccc tgttttttat gagtatatga
1401 taaaatttta tgaacattta cacaaaatga aaatggatat ataggaaaaa
1451 tggagcggta tttcatttat ctttgattgt catttggata ttatattacc
1501 ytgggtaggc aattaaaaat gttaaataac aatttaagga aattatattt
1551 tatatattaa aattaacact gtattatatg attcgcttat aaaagccact
1601 ctttccccat gcaaagctgt ttaatatcaa ttttaacaaa ttacacacat
1651 gttaatatat ttatatatat aatttatata tttataattt atatatttat
1701 atttttatta tttatatatt tattatttat tgtgtgtgtc aattcgggta
1751 ggatatacct cttttttatt gtttaaagcg atttgtattc taaaatataa
1801 agratttgaa aaagagaaag atagaatatg atcccatcat atatagccct
1851 ataattttta tttagcagcg aattaatttt tctattaagt ttatgtgtaa
1901 ttaaaataac ggaatatata taatacaata aaaaagtgca taaattaaaa
1951 ttttttcaat taaatttttt tttttaaggg gttatataat attaaatata
2001 taaaatacga ttatatattt ttgctacaat tttttatatt aagatataaa
2051 tagtaaataa atggtattat atggcatgta atatataaat tttttccaat
2101 ttttatttta tatacacttt tccttttttt gtcataaaac ttaaacaatt
2151 tacacattca ttttaaaaat tgactatttg tttcaacatt ttttgagttt
2201 ccgttttata atagtatttt catttgtata ttgcttatat atataaatac
2251 acacctaaat gttacaaagg atcaatgcat aaaccggtgt gtctggtcgt
2301 cgcgatgacc cccaagaggg gcatcggcat caacaacggc ctcccgtggc
2351 cccacttgac cacagatttc aaacactttt ctcgtgtgac aaaaacgacg
2401 cccgaagaag ccagtcgcct gaacgggtgg cttcccagga aatttgcaaa
2451 gacgggcgac tctggacttc cctctccatc agtcggcaag agattcaacg
2501 ccgttgtcat gggacggaaa acctgggaaa gcatgcctcg aaagtttaga
2551 cccctcgtgg acagattgaa catcgtcgtt tcctcttccc tcaaagaaga
2601 agacattgcg gcggagaagc ctcaagctga aggccagcag cgcgtccgag
2651 tctgtgcttc actcccagca gctctcagcc ttctggagga agagtacaag
2701 gattctgtcg accagatttt tgtcgtggga ggagcgggac tgtacgaggc
2751 agcgctgtct ctgggcgttg cctctcacct gtacatcacg cgtgtagccc
2801 gcgagtttcc gtgcgacgtt ttcttccctg cgttccccgg agatgacatt
2851 ctttcaaaca aatcaactgc tgcgcaggct gcagctcctg ccgagtctgt
2901 gttcgttccc ttttgtccgg agctcggaag agagaaggac aatgaagcga
2951 cgtatcgacc catcttcatt tccaagacct tctcagacaa cggggttccc
3001 tacgactttg tggttctcga gaagagaagg aagactgacg acgcagccac
3051 tgcggaaccg agcaacgcaa tgagctcctt gacgtccacg agggagacaa
3101 ctcccgtgca cgggttgcag gctccttctt cggccgcagc cattgccccg
3151 gtgttggcgt ggatggacga agaagaccgg aaaaaacgcg agcaaaagga
3201 actgattcgg gccgttccgc atgttcactt tagaggccat gaagagttcc
3251 agtaccttga tctcattgcc gacattatta acaatggaag gacaatggat
3301 gaccgaacgg gcgttggtgt catctccaaa ttcggctgca ctatgcgcta
3351 ctcgctggat caggcctttc cacttctcac cacaaagcgt gtgttctgga
3401 aaggggtcct cgaagagttg ctgtggttca ttcgcggcga cacgaacgca
3451 aaccatcttt ctgagaaggg cgtgaagatc tgggacaaga atgtgacacg
3501 cgagttcctc gattcgcgca atctccccca ccgagaggtc ggagacatcg
3551 gcccgggcta cggcttccag tggagacact tcggcgcggc atacaaagac
3601 atgcacacag actacacagg gcagggcgtc gaccagctga agaatgtgat
3651 ccagatgctg agaacgaatc caacagatcg tcgcatgctc atgactgcct
3701 ggaatcctgc agcgctggac gaaatggcgc tgccgccttg tcacttgttg
3751 tgccagttct acgtgaacga ccagaaggag ctgtcgtgca tcatgtatca
3801 gcggtcgtgc gatgtcggcc tcggcgtccc cttcaacatc gcttcctatt
3851 cgcttttgac gctcatggtt gcacacgtct gcaacctaaa acctaaggag
3901 ttcattcact tcatggggaa cacgcatgtc tacacgaacc atgtcgaggc
3951 tttaaaagag cagctgcgga gagaaccgag accgttcccc attgtgaaca
4001 tcctcaacaa ggaacgcatc aaggaaatcg acgatttcac cgccgaggat
4051 tttgaggtcg tgggctacgt cccgcacgga cgaatccaga tggagatggc
4101 tgtctagcgg aaatacagaa gctagctttg atcccgtttt tcttacttat
4151 atatttatac caattgattg tatttataac tgtaaaaatg tgtatgttgt
4201 gtgcatattt ttttttgtgc atgcacatgc atgtaaatag ctaaaattat
4251 gaacatttta ttttttgttc agaaaaaaaa aactttacac acataaaatg
4301 gctagtatga atagccatat tttatataaa ttaaatccta tgaatttatg
4351 accatattaa aaatttagat atttatggaa cataatatgt ttgaaacaat
4401 aagacaaaat tattattatt attattattt ttactgttat aattatgttg
4451 tctcttcaat gattcataaa tagttggact tgatttttaa aatgtttata
4501 atatgattag catagttaaa taaaaaaagt tgaaaaatta aaaaaaaaca
4551 tataaacaca aatgatgttt tttccttcaa tttcgatatc cgcatatcga
4601 aatgatgcta tcatttttta tggagtattg aatatgtgat aaacacaagt
4651 tgagaggggc gtgccgaaat taaataaaat cctctcaaaa ttgatgagaa
4701 gtgtattata ctatgcacat gcttatttac gagtacatgc ctaagcgcta
4751 tttgctaaat tttgtgattt ttcataagaa aatttatagt ttgtgtgtgg
4801 agaatattac taaaatacat agaaagctga atgagaaaac atggaagaaa
4851 aataaaaaaa agaagataaa tagcaataaa aaatattctc taaacacatc
4901 gattttaact acaatattaa aagggtattt aattaattta ttttttgttt
4951 taagtattcg aattcagtat tgcacgacta gatatttgtt ttatattttt
5001 tcaaatagca aattatttta tttctcctta aattgatata tggaaaaaaa
5051 tatataaata aaaaataaca gcaataataa taaacatttg ccttacataa
5101 tagtaatatg caatatatag ctatatgtaa tggtgagtga aaatcagtta
5151 aatgtggggt aaaaaagagt gtatatacct tttttttaaa aagaaaagtt
5201 ttatctgtgc aaaaaaaaaa attaattaaa attattaata tatgtgaatg
5251 ttaagcatat ggatttttta ttaaaggaat atacatacac aataggatga
5301 aatataaaat agtgaaaaaa aaattatatt gtagatggtg tatattctat
5351 tatggcttgt aacgaatatt caaaaaaagt tcatgatgat ttgaatacaa
5401 atccaaacac agaaataatt gcatgcacac gttcatgtac ttgtttaaac
5451 gatgaaatgc gaaaaatgat aatatacatt ttctgtaaaa taaaagaata
5501 cgctacacac tactggatcc ctaaatatat aataatattg ctacaattag
5551 gaatataaat gttttagcca aaaatccata atatgctgaa tctatagtaa
5601 ctgtcatata agcacttttt ttatccttct tacatataca agtaaaagat
5651 gtcgtttttg gtatacttcc aactatacca aataatttaa ttttatcatc
5701 tccttcagca tcttcataat attcaatatc tcctatattt atttgtgaat
5751 ctaaataaac aatgttggat ggttcaagtt cttctgattc aggttgatat
5801 acttgaaaaa agcaatctgg tataatatca ccaggacaat ttaaacctac
5851 caaatgatta tattttggtt cagacaaatg tacgttacat gaaatatgtg
5901 cactatcatc aactaaagaa atatctaaac tatctgtaaa agtatgttta
5951 gaactaacat ttgaagagaa gttacatccg tgtatgactt ttttttcata
6001 ttttggtttt aaaactattt tataattatt atttttaaat ttgcatgtac
6051 attctgtatt aacatctttt gatgtaacat aaaatggagc tttaaagata
6101 gttaactttt catgataaat tattttatta cttttatata aggctttttc
6151 ttttccttct tgaaaacatt ttttattaat tagttcacaa gctaaaacaa
6201 ataattctcc atcattcaat tctacttcta atacattact tacaaaatta
6251 gattcattat atgttttcgg caaatatgta aaaagatcat ttgttaaatt
6301 agtaaataaa atattatgtg gatattttaa tacacgtaca tgtaccatag
6351 cacttctccc ttctatactt gatataacct tttcagtatt atcacaaaaa
6401 caaaaaaatt caatgtcttt tggactaaat ggagatatag ttattgtcct
6451 ttcattataa ttagggtttg tatcattttc ttctatttca tattcaatta
6501 aacctaattc cgatatatca gtttcaactc tattctcata atcagtatat
6551 actttttgaa aacatttttc tggtaatatt ttaaactcag gagacgaact
6601 ttttttaggt attattaatc ttatcttatc atatataaaa tacgaatgga
6651 tggtgcaaaa aatagaccta cgttcacgca tatctggctt taaattatga
6701 acaccttcat ttgaaaaatt acacttatat cctatgaatc cagatatttc
6751 actattcaaa gagctaggct tacaaaaatc attgtttcca cttattattc
6801 ttaatactaa tactatatat aagataaaag caacttgagc ctttttcgca
6851 gaaatatata acatcattct agatattaaa gagagaaaag ggacacacac
6901 ggcaaaaatg cagaaaaaaa ttactcttct tttacttcac tacatatatt
6951 cactgattta tttgcacttg ataactttga tgtgcgggga agtgtgcgtg
7001 ggtattgacg tatgcttata tatgccttct tcgttttatc taagcttaag
7051 caatattata cccatagttt tgtatgattt tcgattcgtt caaaatacca
7101 tacacatgta tgtatgaatg gatatttatt gaataaaaat atctttgtga
7151 atttcgaaaa aaaaataatg catattttga gaaatatttt tttatgtcta
7201 ttattccaat aggctttccg cgaaaaattg gatcaataca tgcacaaaaa
7251 tatataataa catgcgcata ttgtagaatg aattcccact attgatagtt
7301 ttatattatc agttcatagc aatacatgta aataaatata tatataatat
7351 aactaaaaaa atgatgatgg atgaagaatg accgtatttc ttacttaaca
7401 tgcattaaac gatgcgcata taaattcata aaaaatatat tacctacatt
7451 tcatacctaa attaatatta ttatataaac atttattata ttattgtgat
7501 gcgatataat tttaattaaa aaataaaaat atttcatatc tcaataattt
7551 ttttttgaag aatttaaaaa tacaagaaat aaaagcttgg gagtaacata
7601 tatgtttttt tccccttttt atatctattt gaatgactac tattattttt
7651 gtctaatttt atcattagcg ttatgtggat tttggatatt actttcaaaa
7701 gtacactaat ttttattttt taatttattt agttcatatc tacatttttt
7751 ttattatatt tgaaaatatt taatacgatt attagagtaa ttggatttag
7801 aatgtattaa cacacacaca catatatata cggctatatg gaggtatttc
7851 cgcggtggag ctccaattcg ccctatagtg agtcgtatta cgcgcgctca
7901 ctggccgtcg ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca
7951 acttaatcgc cttgcagcac atcccccttt cgccagctgg cgtaatagcg
8001 aagaggcccg caccgatcgc ccttcccaac agttgcgcag cctgaatggc
8051 gaatgggacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt
8101 tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt
8151 tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa
8201 gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca
8251 cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat
8301 cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt
8351 aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt
8401 ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa
8451 aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta
8501 acgcttacaa tttaggtggc acttttcggg gaaatgtgcg cggaacccct
8551 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
8601 ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta
8651 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt
8701 cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga
8751 tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta
8801 agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact
8851 tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca
8901 agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt
8951 actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa
9001 ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact
9051 tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca
9101 tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa
9151 gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac
9201 aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc
9251 aacaattaat agactggatg gaggcggata aagttgcagg accacttctg
9301 cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg
9351 tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc
9401 cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat
9451 gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg
9501 gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac
9551 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
9601 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
9651 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
9701 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
9751 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
9801 agcgcagata ccaaatactg tccttctagt gtagccgtag ttaggccacc
9851 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
9901 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
9951 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
10001 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
10051 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
10101 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
10151 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
10201 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
10251 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
10301 tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct
10351 gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag
10401 ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc
10451 caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc
10501 tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat
10551 taatgtgagt tagctcactc attaggcacc ccaggcttta cactttatgc
10601 ttccggctcg tatgttgtgt ggaattgtga gcggataaca atttcacaca
10651 ggaaacagct atgaccatga ttacgccaag ctcgaaatta accctcacta
10701 aagggaacaa aagctggtac cttctaaaca atcgcagatc gacatattta
10751 attttagaaa agttacatat ctatttgctt catattcttt cttttttttt
10801 attgacagaa ataagaaaaa gggaaattcg aaaattattt attttcttac
10851 aggcaatata agctatccgt ctctgaacat gcgtgatatc attgttgttt
10901 cgtttttttt ttttaatttt tcactatttg cttcatagta tatttcatta
10951 cattttttcc catttttttt aaggcactag gcctcttctc caaacatatc
11001 ttatcaaaag aaaaaatcat gcatgtattt ccagttaaat gcatattttt
11051 gtcttaacaa tgctaatcct tttcctagta ctaaattaat ttgatattta
11101 tacgcaggat aagaagacaa taattcttca tatttgcttt ttaaaaattc
11151 gatatgttca tttatatttt gtattttaaa ggacgctaaa acatgcgttc
11201 ctgctaaacc agtagatcga gatgattgtt ctaatatttc attaatatga
11251 tcacgtaaat cttccaatat aaataaaaca tcaaataatt cattaatttt
11301 ttttttttga tcgattatag acttatcatt ttcataccct ttgatttttt
11351 cttctatatc atttaatcgt ccttcccaaa ttggaacatt taaaaccaaa
11401 tttactgagt cgttccaatt gcttgttatg atattgttgt acaaattttc
11451 catatctccc tttaactctt ctttgccttt cacattgttg tataaatatg
11501 taaagtagtt tcttttattg ccaggaaaaa aaactttctt cgtattaatt
11551 tgatgactat taatgttgcc acaattatta attttgtcta ttaagtaaca
11601 gtatccccta ttttcttgaa tatatttatg tcccttacta taaggtgtat
11651 tttttactct cttgagatgc gcatatcgaa atgatgctat ca"
Restriction sites to linearize plasmid BamH1, KpnI
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 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 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
Sequence Primer 5
Additional information primer 5
Sequence Primer 6
Additional information primer 6