Summary

RMgm-283
Malaria parasiteP. berghei
Genotype
DisruptedGene model (rodent): PBANKA_1432200; Gene model (P.falciparum): PF3D7_1216500; Gene product: male development gene 1 | protein of early gametocyte 3 (MDV-1; PEG3; MDV-1/PEG3)
Phenotype Gametocyte/Gamete; Fertilization and ookinete;
Last modified: 27 May 2009, 09:12
  *RMgm-283
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) : 19438517
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. Ponzi, C.J. Janse, P. Alano
Name Group/DepartmentDipartimento di Malattie Infettive, Parassitarie e Immunomediate
Name InstituteIstituto Superiore di Sanità
CityRome
CountryItaly
Name of the mutant parasite
RMgm numberRMgm-283
Principal name678cl1; 721cl1
Alternative nameΔmdv-1/peg3
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. Both male and female gametocytes showed a strong reduction in egress from the host erythrocyte, resulting in strongly decreased fertilisation rates.
Fertilization and ookineteGametocytes were strongly impaired in their capacity to produce mature ookinetes. Although small numbers of ookinetes were formed, they represented at most 6% (range 0–6%) of the ookinete production observed in wild type parasites. The small number of ookinetes produced normal oocysts and infectious sporozoites.
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 MDV-1/PEG3.

Protein (function)
The protein is a gametocyte specific protein and has been named in P. falciparum MDV-1 (male development gene 1) or PEG3. In P. falciparum gametocytes the protein is associated with the osmiophilic bodies, the parasite membrane and all membranous compartments derived from the parasitophorous vacuole membrane. P. falciparum parasites lacking expression of MDV-1/PEG3 were affected in their capacity to produce functional gametocytes, which were unable to be transmitted through mosquitoes.

Phenotype
The phenotype analyses indicate that the lack of expression of MDV-1/PEG3 strongly impairs fertilisation and ookinete formation. The phenotype analyses indicate that the decreased fertilisation rate results from a strongly reduced capacity of both male and female gametocytes/gametes to egress from their host erythrocytes (see also 'Additional information'). The results indicate that MDV-1/PEG3 plays a role in disruption of the parasitophorous vacuole membrane (PVM) and the erythrocyte membrane.

Additional information
MDV-1/PEG3 is expressed in both male and female gametocytes with higher expression in females compared to males. Immunoelectron microcope analysis showed an association of MDV-1/PEG3 with the osmiophilic bodies in female gametocytes. Evidence is presented that during female gamete formation MDV-1/PEG3 is released extracellularly. 
Normal numbers of male and female gametocytes are produced that are able to form gametes.
Cross-fertilisation studies with wild type male and female gametes showed that both mutant female and mutant male gametocytes/gametes are affected in fertilisation and ookinete formation.
Live light microscope observations on gamete formation and electron microscope analysis provided evidence that both male and female gametes are strongly impaired in emergence/egress from their host erythrocytes and failed to disrupt the parasitophorous vacuole membrane (PVM) and the erythrocyte membrane.
A second mutant lacking expression of MDV-1/PEG3 has been generated in the same study (RMgm-282). This mutant expresses GFP in male gametocytes/gametes and RFP in female gametocytes/gametes/zygotes.

An independent mutant lacking expression of MDV-1/PEG3 has been described (RMgm-231) that showed a slightly different phenotype. Both female and male gametocytes were strongly impaired in fertilisation and production of ookinetes. In addition, ookinetes lacking expression of MDV-1/PEG3 showed an impaired development, suggesting of an additional role of MDV-1/PEG3 in the ookinete stage.

Other mutants
RMgm-282: A mutant lacking expression of MDV-1/PEG3, generated in the same study. This mutant expresses GFP in male gametocytes/gametes and RFP in female gametocytes/gametes/zygotes.
RMgm-231: An independent mutant lacking expression of MDV-1/PEG3.
RMgm-251: A mutant expressing GFP under the control of the MDV-1/PEG3 promoter.


  Disrupted: Mutant parasite with a disrupted gene
Details of the target gene
Gene Model of Rodent Parasite PBANKA_1432200
Gene Model P. falciparum ortholog PF3D7_1216500
Gene productmale development gene 1 | protein of early gametocyte 3
Gene product: Alternative nameMDV-1; PEG3; MDV-1/PEG3
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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ATCCCCTGAAGAAGAAATTGATCATTTTGCAGATGATTTAACTGATAAATATGAAGTTAA
GCAAAATGAAAATTTAGAACAATATGAAAAAGACATAAATCCACATCATTAAAATTAATA
TGGAATAATAACTAAACATGCCCGTGTATTTGCGCCAATATATGCGCATACATATGTATA
TATGCATATTCCAAATGATCAAGGACATTTTTTGTTGTTGAACTAAACTTTATAAGTATG
ATTTAAATGTATAGAAACAAAATTTGTAATAAGAAACTATTCATAACTTCATAATAATTA
TAAACATATGAATTGTGTTTTTTCTTTCATTTATTTTATAAATGCCATTTATATTATCGT
AGTTAATTTAGAAAATTTTTTTTTTTCAAATTTCGTGATCGTCATTATGAGTATAGTTGT
TTTATTATTTTTACATTCATTATATTTTACGTGTGTATGGCGCATCTATTATTCTTTACA
TATTGAGTAAATAAAAATTCTTTTATATGCGTATCCATATTTTTTTCCTAATCTTATTTA
TTTATACATTTTTGCATAAGCTTGTGCACGAAAATCCACATAAAATATCTATAAATATAG
ACATATTCAAAACACACCAAATGTAAAAAATAACTGAGAGGAATTTAACAAATAGAATAG
TTTATACAATCGTTTTTTTCCATTTATGGGAAAAGTGTACACTATTAACATGCTAATGTT
GTAAAAGTTATGGATTAAAAAAATTAAAATCGACACTGTTAGACTAAGTTAGAATTATAT
GCGGATCCACTAGTTCTAGAGCGGCCGCCACCGCGGTGGAGCTCCAATTCGCCCTATAGT
GAGTCGTATTACGCGCGCTCACTGGCCGTCGTTTTACAACGTCGTGACTGGGAAAACCCT
GGCGTTACCCAACTTAATCGCCTTGCAGCACATCCCCCTTTCGCCAGCTGGCGTAATAGC
GAAGAGGCCCGCACCGATCGCCCTTCCCAACAGTTGCGCAGCCTGAATGGCGAATGGGAC
GCGCCCTGTAGCGGCGCATTAAGCGCGGCGGGTGTGGTGGTTACGCGCAGCGTGACCGCT
ACACTTGCCAGCGCCCTAGCGCCCGCTCCTTTCGCTTTCTTCCCTTCCTTTCTCGCCACG
TTCGCCGGCTTTCCCCGTCAAGCTCTAAATCGGGGGCTCCCTTTAGGGTTCCGATTTAGT
GCTTTACGGCACCTCGACCCCAAAAAACTTGATTAGGGTGATGGTTCACGTAGTGGGCCA
TCGCCCTGATAGACGGTTTTTCGCCCTTTGACGTTGGAGTCCACGTTCTTTAATAGTGGA
CTCTTGTTCCAAACTGGAACAACACTCAACCCTATCTCGGTCTATTCTTTTGATTTATAA
GGGATTTTGCCGATTTCGGCCTATTGGTTAAAAAATGAGCTGATTTAACAAAAATTTAAC
GCGAATTTTAACAAAATATTAACGCTTACAATTTAGGTGGCACTTTTCGGGGAAATGTGC
GCGGAACCCCTATTTGTTTATTTTTCTAAATACATTCAAATATGTATCCGCTCATGAGAC
AATAACCCTGATAAATGCTTCAATAATATTGAAAAAGGAAGAGTATGAGTATTCAACATT
TCCGTGTCGCCCTTATTCCCTTTTTTGCGGCATTTTGCCTTCCTGTTTTTGCTCACCCAG
AAACGCTGGTGAAAGTAAAAGATGCTGAAGATCAGTTGGGTGCACGAGTGGGTTACATCG
AACTGGATCTCAACAGCGGTAAGATCCTTGAGAGTTTTCGCCCCGAAGAACGTTTTCCAA
TGATGAGCACTTTTAAAGTTCTGCTATGTGGCGCGGTATTATCCCGTATTGACGCCGGGC
AAGAGCAACTCGGTCGCCGCATACACTATTCTCAGAATGACTTGGTTGAGTACTCACCAG
TCACAGAAAAGCATCTTACGGATGGCATGACAGTAAGAGAATTATGCAGTGCTGCCATAA
CCATGAGTGATAACACTGCGGCCAACTTACTTCTGACAACGATCGGAGGACCGAAGGAGC
TAACCGCTTTTTTGCACAACATGGGGGATCATGTAACTCGCCTTGATCGTTGGGAACCGG
AGCTGAATGAAGCCATACCAAACGACGAGCGTGACACCACGATGCCTGTAGCAATGGCAA
CAACGTTGCGCAAACTATTAACTGGCGAACTACTTACTCTAGCTTCCCGGCAACAATTAA
TAGACTGGATGGAGGCGGATAAAGTTGCAGGACCACTTCTGCGCTCGGCCCTTCCGGCTG
GCTGGTTTATTGCTGATAAATCTGGAGCCGGTGAGCGTGGGTCTCGCGGTATCATTGCAG
CACTGGGGCCAGATGGTAAGCCCTCCCGTATCGTAGTTATCTACACGACGGGGAGTCAGG
CAACTATGGATGAACGAAATAGACAGATCGCTGAGATAGGTGCCTCACTGATTAAGCATT
GGTAACTGTCAGACCAAGTTTACTCATATATACTTTAGATTGATTTAAAACTTCATTTTT
AATTTAAAAGGATCTAGGTGAAGATCCTTTTTGATAATCTCATGACCAAAATCCCTTAAC
GTGAGTTTTCGTTCCACTGAGCGTCAGACCCCGTAGAAAAGATCAAAGGATCTTCTTGAG
ATCCTTTTTTTCTGCGCGTAATCTGCTGCTTGCAAACAAAAAAACCACCGCTACCAGCGG
TGGTTTGTTTGCCGGATCAAGAGCTACCAACTCTTTTTCCGAAGGTAACTGGCTTCAGCA
GAGCGCAGATACCAAATACTGTCCTTCTAGTGTAGCCGTAGTTAGGCCACCACTTCAAGA
ACTCTGTAGCACCGCCTACATACCTCGCTCTGCTAATCCTGTTACCAGTGGCTGCTGCCA
GTGGCGATAAGTCGTGTCTTACCGGGTTGGACTCAAGACGATAGTTACCGGATAAGGCGC
AGCGGTCGGGCTGAACGGGGGGTTCGTGCACACAGCCCAGCTTGGAGCGAACGACCTACA
CCGAACTGAGATACCTACAGCGTGAGCTATGAGAAAGCGCCACGCTTCCCGAAGGGAGAA
AGGCGGACAGGTATCCGGTAAGCGGCAGGGTCGGAACAGGAGAGCGCACGAGGGAGCTTC
CAGGGGGAAACGCCTGGTATCTTTATAGTCCTGTCGGGTTTCGCCACCTCTGACTTGAGC
GTCGATTTTTGTGATGCTCGTCAGGGGGGCGGAGCCTATGGAAAAACGCCAGCAACGCGG
CCTTTTTACGGTTCCTGGCCTTTTGCTGGCCTTTTGCTCACATGTTCTTTCCTGCGTTAT
CCCCTGATTCTGTGGATAACCGTATTACCGCCTTTGAGTGAGCTGATACCGCTCGCCGCA
GCCGAACGACCGAGCGCAGCGAGTCAGTGAGCGAGGAAGCGGAAGAGCGCCCAATACGCA
AACCGCCTCTCCCCGCGCGTTGGCCGATTCATTAATGCAGCTGGCACGACAGGTTTCCCG
ACTGGAAAGCGGGCAGTGAGCGCAACGCAATTAATGTGAGTTAGCTCACTCATTAGGCAC
CCCAGGCTTTACACTTTATGCTTCCGGCTCGTATGTTGTGTGGAATTGTGAGCGGATAAC
AATTTCACACAGGAAACAGCTATGACCATGATTACGCCAAGCTCGAAATTAACCCTCACT
AAAGGGAACAAAAGCTGGTACCCAAATAATGGCCAATTTAATTAATAACGGTATTATAGT
TAAAATAAAGTTCATGCTAATTTATTAATTATATTTACAAAATAAGGAAATATGTGTATT
AGACAGCAATTTCTATGTATTTTTTATATACTATTAAAAGCTCAATTGCTCAAAAAATAA
ATAAGTATTTCTTCTATAAAAAATATCATCGAAAAAAATATATAAGATTACATTTAAAAA
AACTTATTTACATTGGGTATTTTAGGCAAAACGGGATATATATTCAGCTCAATGCACATC
TAGTTATTACATTAAATATGAAAAAAAAAAAACAATTTCATCTCAATCTAGCTAATACTT
AACTAATATTTTTAAAATAAAGAAAATACACCTTGCACATAAAAAAGAAGAAAAATTAAA
AAAAAATTGTATATGTGCACACTATATTTATTAATTTTTTTTATCATAAAATATATAAAT
TGTGAAAATACACTAAAAATTGTAGATTTTAGTAAAAATTACTTACGGATTAGATATGTG
CAAAATACGAAAAATTAGCAAATGCAAAATAAATATAAAATTAATATCCATACTATATAT
ATTTTAAGACAAATTTTTTTAAAACCAGCTTTTTTAAAATCGTAATTAGTGATATATAAT
TTAATGATATAAAAATATTTTTTATGTTTTACATATTTATGAGGCAATTATTTTCATTAG
GGCTATCGTTTAATATATAAAATTTTAAAAATTGAAATAAACAAAATAAGTTATATAATC
GTATAAGAAAATATATATTTATAAAGATTAAACTATTTACATTATAATAATTAGTAAAAA
AAAAATATATACTATTCACAATGAAGTGTATTAATATTCCAACATCAACCATAGGGTGTC
TATTATTATTTTTTTTGAGCTCACAAAATGCTCAATACGAGTGCATGAATATAAAAGGTT
CATCAAATAATGGAAGTGGAGATCGATTCTAAAGGGCGAATTCCAGCACACTGGCGGCCG
TTACTAGTGGATCCGAGCTCGGTACCGGGCCCCCCCTCGAGGTCGACGGTATCGATAAGC
TTGCATGCCTGCAGGTCAACAATAAATAATAAATAAATATTGTGGAAATAAAATAACATA
TAATTATTTTTAATACATTGATTTCCCTTTTATTTTTTTAAATTTCATTGATATAAAAAT
ATATAATAATAACATATATGATTTCAAATTAATCTTTTCAAAAATGGTGTTTATTTTTGT
ATGTTTGTGTATGAATTAATCACATAACACATCTATTAAATTGAGTTGGTAATATAGACA
CAAATAAATATATATATTTTTATAGCTTAAAAGTGTGTTATGAATATTTTAAGCATATTT
TCTTTTTCTTTGGATTGTGTAAAATGAACTCATATAATGCGTTTTTTTGTTTTTGTTATT
TTGTCATTTTGTTATTTTGCTATTTTATGGATTAATTTTTGTTTATAAAATGGGAAATAA
TTTTAACATATTTAAATAAATGGAGAAAAAATATAAAATAATTATAAAAAAAAGTTAATA
CACATTTTTTCCTGTTATAGACCTTATATTTATTTATCCATATATATATATATATATATA
TATATATATATATACATACCAAGTGAATTAAGAGGAAAGCTAATTTATTATTCAGAATAA
TATATGAACTATATATAATTTTTATTATTTTGGTGTATATTAATCTGTCTATATGCATAC
ATGCAATAATTTATCGACTTATATATCAAATAACATAAAATAGAAGTGTTTTAAATTATG
GATATATGCTCAATATTCATTTTTTTTAATAAGTTAGCTATATTTAAATTATACATTTTA
TATATGGTCTCTTTTTTTTTTAAATATTATTTAAGTGATCATGAAAATATAAATAATTTT
TTTTTATTTAATATCCTTTTGCTTGCATGTGGTAAATGGAAATTTGGATGTGTTTTGAAR
GTTCGGATATAGTTGTATGGACATATAATATATTTTGTGAAAAATTGGTTTTATGTTTAT
ACTTATGCCAATACTTTTTGAGTAAAACAAAGCAAGTGCTTATAAATAATTAAAGCCAAT
TTTATAATATATATTTTTTTATTTAATTTGAATTTAGTAGTATAATTTTTTATGGTAAGT
GCTCAAAGAGAGTTGCTTATAAAGTATGGTTTGTTTCTTTTTCGCCATTTTGAATTACAC
ATTAAAAATATATAGATACATATATTATAATATGAAATCATTAATAATTTAGGGAAATTC
TACAAATTTAAAAACGAATAAAATAATTGTTTTTCATCATGCCATAACACAATATTGATA
TATACATGTACAAACATTTTTTTTATTTGGAAAATATAAATTATATAAAAAAAAATGTAT
AGTATACAAAATGAGCATATTCACACGGGGTGGACGTTCATTTTTTCATTTTTCCCCTGT
TTTTTATGAGTATATGATAAAATTTTATGAACATTTACACAAAATGAAAATGGATATATA
GGAAAAATGGAGCGGTATTTCATTTATCTTTGATTGTCATTTGGATATTATATTACCYTG
GGTAGGCAATTAAAAATGTTAAATAACAATTTAAGGAAATTATATTTTATATATTAAAAT
TAACACTGTATTATATGATTCGCTTATAAAAGCCACTCTTTCCCCATGCAAAGCTGTTTA
ATATCAATTTTAACAAATTACACACATGTTAATATATTTATATATATAATTTATATATTT
ATAATTTATATATTTATATTTTTATTATTTATATATTTATTATTTATTGTGTGTGTCAAT
TCGGGTAGGATATACCTCTTTTTTATTGTTTAAAGCGATTTGTATTCTAAAATATAAAGR
ATTTGAAAAAGAGAAAGATAGAATATGATCCCATCATATATAGCCCTATAATTTTTATTT
AGCAGCGAATTAATTTTTCTATTAAGTTTATGTGTAATTAAAATAACGGAATATATATAA
TACAATAAAAAAGTGCATAAATTAAAATTTTTTCAATTAAATTTTTTTTTTTAAGGGGTT
ATATAATATTAAATATATAAAATACGATTATATATTTTTGCTACAATTTTTTATATTAAG
ATATAAATAGTAAATAAATGGTATTATATGGCATGTAATATATAAATTTTTTCCAATTTT
TATTTTATATACACTTTTCCTTTTTTTGTCATAAAACTTAAACAATTTACACATTCATTT
TAAAAATTGACTATTTGTTTCAACATTTTTTGAGTTTCCGTTTTATAATAGTATTTTCAT
TTGTATATTGCTTATATATATAAATACACACCTAAATGTTACAAAGGATCAATGCATAAA
CCGGTGTGTCTGGTCGTCGCGATGACCCCCAAGAGGGGCATCGGCATCAACAACGGCCTC
CCGTGGCCCCACTTGACCACAGATTTCAAACACTTTTCTCGTGTGACAAAAACGACGCCC
GAAGAAGCCAGTCGCCTGAACGGGTGGCTTCCCAGGAAATTTGCAAAGACGGGCGACTCT
GGACTTCCCTCTCCATCAGTCGGCAAGAGATTCAACGCCGTTGTCATGGGACGGAAAACC
TGGGAAAGCATGCCTCGAAAGTTTAGACCCCTCGTGGACAGATTGAACATCGTCGTTTCC
TCTTCCCTCAAAGAAGAAGACATTGCGGCGGAGAAGCCTCAAGCTGAAGGCCAGCAGCGC
GTCCGAGTCTGTGCTTCACTCCCAGCAGCTCTCAGCCTTCTGGAGGAAGAGTACAAGGAT
TCTGTCGACCAGATTTTTGTCGTGGGAGGAGCGGGACTGTACGAGGCAGCGCTGTCTCTG
GGCGTTGCCTCTCACCTGTACATCACGCGTGTAGCCCGCGAGTTTCCGTGCGACGTTTTC
TTCCCTGCGTTCCCCGGAGATGACATTCTTTCAAACAAATCAACTGCTGCGCAGGCTGCA
GCTCCTGCCGAGTCTGTGTTCGTTCCCTTTTGTCCGGAGCTCGGAAGAGAGAAGGACAAT
GAAGCGACGTATCGACCCATCTTCATTTCCAAGACCTTCTCAGACAACGGGGTTCCCTAC
GACTTTGTGGTTCTCGAGAAGAGAAGGAAGACTGACGACGCAGCCACTGCGGAACCGAGC
AACGCAATGAGCTCCTTGACGTCCACGAGGGAGACAACTCCCGTGCACGGGTTGCAGGCT
CCTTCTTCGGCCGCAGCCATTGCCCCGGTGTTGGCGTGGATGGACGAAGAAGACCGGAAA
AAACGCGAGCAAAAGGAACTGATTCGGGCCGTTCCGCATGTTCACTTTAGAGGCCATGAA
GAGTTCCAGTACCTTGATCTCATTGCCGACATTATTAACAATGGAAGGACAATGGATGAC
CGAACGGGCGTTGGTGTCATCTCCAAATTCGGCTGCACTATGCGCTACTCGCTGGATCAG
GCCTTTCCACTTCTCACCACAAAGCGTGTGTTCTGGAAAGGGGTCCTCGAAGAGTTGCTG
TGGTTCATTCGCGGCGACACGAACGCAAACCATCTTTCTGAGAAGGGCGTGAAGATCTGG
GACAAGAATGTGACACGCGAGTTCCTCGATTCGCGCAATCTCCCCCACCGAGAGGTCGGA
GACATCGGCCCGGGCTACGGCTTCCAGTGGAGACACTTCGGCGCGGCATACAAAGACATG
CACACAGACTACACAGGGCAGGGCGTCGACCAGCTGAAGAATGTGATCCAGATGCTGAGA
ACGAATCCAACAGATCGTCGCATGCTCATGACTGCCTGGAATCCTGCAGCGCTGGACGAA
ATGGCGCTGCCGCCTTGTCACTTGTTGTGCCAGTTCTACGTGAACGACCAGAAGGAGCTG
TCGTGCATCATGTATCAGCGGTCGTGCGATGTCGGCCTCGGCGTCCCCTTCAACATCGCT
TCCTATTCGCTTTTGACGCTCATGGTTGCACACGTCTGCAACCTAAAACCTAAGGAGTTC
ATTCACTTCATGGGGAACACGCATGTCTACACGAACCATGTCGAGGCTTTAAAAGAGCAG
CTGCGGAGAGAACCGAGACCGTTCCCCATTGTGAACATCCTCAACAAGGAACGCATCAAG
GAAATCGACGATTTCACCGCCGAGGATTTTGAGGTCGTGGGCTACGTCCCGCACGGACGA
ATCCAGATGGAGATGGCTGTCTAGCGGAAATACAGAAGCTAGCTTTGATCCCGTTTTTCT
TACTTATATATTTATACCAATTGATTGTATTTATAACTGTAAAAATGTGTATGTTGTGTG
CATATTTTTTTTTGTGCATGCACATGCATGTAAATAGCTAAAATTATGAACATTTTATTT
TTTGTTCAGAAAAAAAAAACTTTACACACATAAAATGGCTAGTATGAATAGCCATATTTT
ATATAAATTAAATCCTATGAATTTATGACCATATTAAAAATTTAGATATTTATGGAACAT
AATATGTTTGAAACAATAAGACAAAATTATTATTATTATTATTATTTTTACTGTTATAAT
TATGTTGTCTCTTCAATGATTCATAAATAGTTGGACTTGATTTTTAAAATGTTTATAATA
TGATTAGCATAGTTAAATAAAAAAAGTTGAAAAATTAAAAAAAAACATATAAACACAAAT
GATGTTTTTTCCTTCAATTTCGAT
Restriction sites to linearize plasmid MluNI/XbaI
Partial or complete disruption of the genePartial
Additional remarks partial/complete disruption First 122 bp of the 645bp ORF intact
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 1TAGAggtaccCAAATAATGGCCAATTTAATTAATAACGG
Additional information primer 1L2156 (Asp718); 5'
Sequence Primer 2TAGAatcgatCTCCACTTCCATTATTTGATGAAC
Additional information primer 2L2157 (ClaI); 5' (ClaI site methylated!)
Sequence Primer 3TAGAgatatcCCCTGAAGAAGAAATTGATCATTTTGCAG
Additional information primer 3L2158 (EcoRV); 3'
Sequence Primer 4TAGAggatccGCATATAATTCTAACTTAGTCTAACAGTGTCG
Additional information primer 4L2159 (BamHI); 3'
Sequence Primer 5
Additional information primer 5
Sequence Primer 6
Additional information primer 6