Summary

RMgm-251
Malaria parasiteP. berghei
Genotype
Transgene
Transgene not Plasmodium: GFP
Promoter: Gene model: 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)
3'UTR: Gene model: PBANKA_0719300; Gene product: bifunctional dihydrofolate reductase-thymidylate synthase, putative (dhfr/ts)
Insertion locus: Gene model: Not available; Gene product: Not available (small subunit ribosomal rna gene (c-type unit))
Phenotype Gametocyte/Gamete;
Last modified: 28 May 2009, 10:19
  *RMgm-251
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) : 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 ed Immunomediate
Name InstituteIstituto Superiore di Sanità
CityRome
CountryItaly
Name of the mutant parasite
RMgm numberRMgm-251
Principal name707cl1
Alternative name
Standardized name
Is the mutant parasite cloned after genetic modificationYes
Phenotype
Asexual blood stageNot different from wild type
Gametocyte/GameteNo GFP expression in asexual blood stages. GFP expression in both male and female gametocytes with higher expression in females.
Fertilization and ookineteNot tested
OocystNot tested
SporozoiteNot tested
Liver stageNot tested
Additional remarks phenotype

Mutant/mutation
The mutant expresses GFP under the control of the MDV-1/PEG3 promoter. The gfp gene is stably integrated into the non-essential c/d-type ssurrna gene.

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.
In P. berghei mutants lacking expression of MDV-1/PEG3 (RMgm-231, RMgm-282, RMgm-283) fertilisation and ookinete formation is strongly impaired. The phenotype analyses of these mutants 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. These results indicate that MDV-1/PEG3 plays a role in disruption of the parasitophorous vacuole membrane (PVM) and the erythrocyte membrane.

Phenotype
No GFP expression in asexual blood stages. GFP expression in both male and female gametocytes with higher expression in females.

Additional information
IFA's using anti-MDV-1/PEG3 antibodies also showed higher expression levels in females compared to males.
The difference in GFP expression between males and female gametocytes allows for separation and counting of the two sexes using FACS (flow cytometry).
Immunoelectron microscope analysis showed an association of MDV-1/PEG3 with the osmiophilic bodies in female gametocytes.

Other mutants
RMgm-231: An independent mutant lacking expression of MDV-1/PEG3.
RMgm-282: A mutant lacking expression of MDV-1/PEG3. This mutant expresses GFP in male gametocytes/gametes and RFP in female gametocytes/gametes/zygotes.
RMgm-283: A mutant lacking expression of MDV-1/PEG3.


  Transgene: Mutant parasite expressing a transgene
Type and details of transgene
Is the transgene Plasmodium derived Transgene: not Plasmodium
Transgene nameGFP
Details of the genetic modification
Inducable system usedNo
Additional remarks inducable system
Type of plasmid/constructPlasmid single 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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TATGCTTGTCTCAAAGATTAAGCCATGCAAGTGAAAGTATATGCACATTTATTGCAGAAA
CTGCGAACGGCTCATTAAAACAGTTATAATCTACTTGACATTTTATTATAAGGATAACTA
CGGAAAAGCTGTAGCTAATACTTGTTAAGTACTTTTACTCCCCGGAGTAATTGTATGTAT
TTGTTAAGACCCCTAAGAAAAAATGATATTAAAGGAATTATAACAAAGAAGCAACACATA
ATATAATTATTCAGTGTGTATCAATCGAGTTTCTGACCTATCAGCTTTTGATGTTAGGGT
ATTGACCTAACATGGCTTTGACGGGTAACGGGGAATTAGAGTTCGATTCCGGAGAGGGAG
CCTGAGAAATAGCTACCACATCTAAGGAAGGCAGCAGGCGCGTAAATTACCCAATTCTAA
ATAAGAGAGGTAGTGACAAGAAATAACAATATAAGGCCAAATTTTGGTTTTATAATTGGA
ATGATGGGAATTTAAAACCTTCCCAAAAATCAATTGGAGGGCAAGTCTGGTGCCAGCAGC
CGCGGTAATTCCAGCTCCAATAGCGTATATTAAAATTGTTGCAGTTAAAACGCTCGTAGT
TGAACTTCAAGGGTATAATTATTTTAAGCAACTCACTTGGAAAGAATCATGACTTCTGTC
ACTGCTTTTATCCTTGTTGCAGTTCTTTTAATACAGGGCCCTTTGAGAGCCCATTAATTT
ATGACTGGGTTTCTCGTTACTTTGAGTAAATTAGAGTGTTTAAAGCAAACAGATAAAGCG
TATTTTACTGTGTTTGAATACTATAGCATGGAATAACAACATTGAATAGGTCAAAAGTTT
TTGAAAAATTTTTCTTATTTTGGCTTAGATACAGTTAATAGGAGTAGCTTGGGGGCATTT
GTATTCAGATGTCAGAGGTGAAATTCTTAGATTTTCTGGAGACAAACAACTGCGAAAGCA
TTTGCCTAAAATACTTCCATTAATCAAGAACGAAAGTTAAGGGAGTGAAGACGATCAGAT
ACCGTCGTAATCTTAACCATAAACTATGCCGACTAAGTGTTGGATGAAAATTTATAAATA
AAACTATCTTCTTTAAAGGAGTAGTTTTTTAGATGCTTCCTTCAGTACCTTATGAGAAAT
CAAAGTCTTTGGGTTCTGGGGCGAGTATTCGCGCAAGCGAGAAAGTTAAAAGAATTGACG
GAAGGGCACCACCAGGCGTGGAGCTTGCGGCTTAATTTGACTCAACACGGGGAAACTCAC
TAGTTTAAGACAAGAGTAGGATTGACAGATTAATAGCTCTTTCTTGATTTCTTGGATGGT
GATGCATGGCCGTTTTTAGTTCGTGAATATGATTTGTCTGGTTAATTCCGATAACGAACG
AGATCTTAACCTGCTAATTAGCGGTGGATATGTGATATTCTTCGAAGGTGGACTAACTAT
AGCGTTTTCGAAGGTATGTTGCATAATCAAATTGGTTTACCCTTTGTTTTTTTGTAGCAT
ATTCTTTTATTTCGTTGGGTTTTTTCCCTAGTAAGGATGTATCTGCTTTATTTAATGCTT
CTTAGAGGAACGATGTGTGTCTAACACAAGGAAGTTTAAGGCAACAACAGGTCTGTGATG
TCCTTAGATATACTAGGCTGCACGCGTGCTACACTGATATGTAAAACGAGTATTTAAAAT
TATATCTGTATGGTAGATAATTTAATTTCTACGTATTATCAGCATATACTTTTCCTACAC
TGAAATAGTGAAGGTAATCTTTATCAATACATATCGTGATGGGGATAGATTATTGCAATT
ATTAATCTTGAACGAGGAATGCCTAGTAAGCATGATTCATCAGATTGTGCTGACTACGTC
CCTGCCCTTTGTACACACCGCCCGTCGCTCCTACCGATTGAAAGATATGATGAATTGTTT
GGACAAGAAAATAGAAATTTTATTTTTATTTTTTTTGGAAGGACCGTAAATCCTATCTTT
TAAAGGAAGGAGAAGTCGTAACAAGGTTTCCGTAGGTGAATTCACTGGCCGTCGTTTTAC
AACGTCGTGACTGGGAAAACCCTGGCGTTACCCAACTTAATCGCCTTGCAGCACATCCCC
CTTTCGCCAGCTGGCGTAATAGCGAAGAGGCCCGCACCGATCGCCCTTCCCAACAGTTGC
GCAGCCTGAATGGCGAATGGCGCCTGATGCGGTATTTTCTCCTTACGCATCTGTGCGGTA
TTTCACACCGCATATGGTGCACTCTCAGTACAATCTGCTCTGATGCCGCATAGTTAAGCC
AGCCCCGACACCCGCCAACACCCGCTGACGCGCCCTGACGGGCTTGTCTGCTCCCGGCAT
CCGCTTACAGACAAGCTGTGACCGTCTCCGGGAGCTGCATGTGTCAGAGGTTTTCACCGT
CATCACCGAAACGCGCGAGACGAAAGGGCCTCGTGATACGCCTATTTTTATAGGTTAATG
TCATGATAATAATGGTTTCTTAGACGTCAGGTGGCACTTTTCGGGGAAATGTGCGCGGAA
CCCCTATTTGTTTATTTTTCTAAATACATTCAAATATGTATCCGCTCATGAGACAATAAC
CCTGATAAATGCTTCAATAATATTGAAAAAGGAAGAGTATGAGTATTCAACATTTCCGTG
TCGCCCTTATTCCCTTTTTTGCGGCATTTTGCCTTCCTGTTTTTGCTCACCCAGAAACGC
TGGTGAAAGTAAAAGATGCTGAAGATCAGTTGGGTGCACGAGTGGGTTACATCGAACTGG
ATCTCAACAGCGGTAAGATCCTTGAGAGTTTTCGCCCCGAAGAACGTTTTCCAATGATGA
GCACTTTTAAAGTTCTGCTATGTGGCGCGGTATTATCCCGTATTGACGCCGGGCAAGAGC
AACTCGGTCGCCGCATACACTATTCTCAGAATGACTTGGTTGAGTACTCACCAGTCACAG
AAAAGCATCTTACGGATGGCATGACAGTAAGAGAATTATGCAGTGCTGCCATAACCATGA
GTGATAACACTGCGGCCAACTTACTTCTGACAACGATCGGAGGACCGAAGGAGCTAACCG
CTTTTTTGCACAACATGGGGGATCATGTAACTCGCCTTGATCGTTGGGAACCGGAGCTGA
ATGAAGCCATACCAAACGACGAGCGTGACACCACGATGCCTGTAGCAATGGCAACAACGT
TGCGCAAACTATTAACTGGCGAACTACTTACTCTAGCTTCCCGGCAACAATTAATAGACT
GGATGGAGGCGGATAAAGTTGCAGGACCACTTCTGCGCTCGGCCCTTCCGGCTGGCTGGT
TTATTGCTGATAAATCTGGAGCCGGTGAGCGTGGGTCTCGCGGTATCATTGCAGCACTGG
GGCCAGATGGTAAGCCCTCCCGTATCGTAGTTATCTACACGACGGGGAGTCAGGCAACTA
TGGATGAACGAAATAGACAGATCGCTGAGATAGGTGCCTCACTGATTAAGCATTGGTAAC
TGTCAGACCAAGTTTACTCATATATACTTTAGATTGATTTAAAACTTCATTTTTAATTTA
AAAGGATCTAGGTGAAGATCCTTTTTGATAATCTCATGACCAAAATCCCTTAACGTGAGT
TTTCGTTCCACTGAGCGTCAGACCCCGTAGAAAAGATCAAAGGATCTTCTTGAGATCCTT
TTTTTCTGCGCGTAATCTGCTGCTTGCAAACAAAAAAACCACCGCTACCAGCGGTGGTTT
GTTTGCCGGATCAAGAGCTACCAACTCTTTTTCCGAAGGTAACTGGCTTCAGCAGAGCGC
AGATACCAAATACTGTCCTTCTAGTGTAGCCGTAGTTAGGCCACCACTTCAAGAACTCTG
TAGCACCGCCTACATACCTCGCTCTGCTAATCCTGTTACCAGTGGCTGCTGCCAGTGGCG
ATAAGTCGTGTCTTACCGGGTTGGACTCAAGACGATAGTTACCGGATAAGGCGCAGCGGT
CGGGCTGAACGGGGGGTTCGTGCACACAGCCCAGCTTGGAGCGAACGACCTACACCGAAC
TGAGATACCTACAGCGTGAGCATTGAGAAAGCGCCACGCTTCCCGAAGGGAGAAAGGCGG
ACAGGTATCCGGTAAGCGGCAGGGTCGGAACAGGAGAGCGCACGAGGGAGCTTCCAGGGG
GAAACGCCTGGTATCTTTATAGTCCTGTCGGGTTTCGCCACCTCTGACTTGAGCGTCGAT
TTTTGTGATGCTCGTCAGGGGGGCGGAGCCTATGGAAAAACGCCAGCAACGCGGCCTTTT
TACGGTTCCTGGCCTTTTGCTGGCCTTTTGCTCACATGTTCTTTCCTGCGTTATCCCCTG
ATTCTGTGGATAACCGTATTACCGCCTTTGAGTGAGCTGATACCGCTCGCCGCAGCCGAA
CGACCGAGCGCAGCGAGTCAGTGAGCGAGGAAGCGGAAGAGCGCCCAATACGCAAACCGC
CTCTCCCCGCGCGTTGGCCGATTCATTAATGCAGCTGGCACGACAGGTTTCCCGACTGGA
AAGCGGGCAGTGAGCGCAACGCAATTAATGTGAGTTAGCTCACTCATTAGGCACCCCAGG
CTTTACACTTTATGCTTCCGGCTCGTATGTTGTGTGGAATTGTGAGCGGATAACAATTTC
ACACAGGAAACAGCTATGACCATGATTACGCCAAGCTTGCATGCCTGCAGGTCAACAATA
AATAATAAATAAATATTGTGGAAATAAAATAACATATAATTATTTTTAATACATTGATTT
CCCTTTTATTTTTTTAAATTTCATTGATATAAAAATATATAATAATAACATATATGATTT
CAAATTAATCTTTTCAAAAATGGTGTTTATTTTTGTATGTTTGTGTATGAATTAATCACA
TAACACATCTATTAAATTGAGTTGGTAATATAGACACAAATAAATATATATATTTTTATA
GCTTAAAAGTGTGTTATGAATATTTTAAGCATATTTTCTTTTTCTTTGGATTGTGTAAAA
TGAACTCATATAATGCGTTTTTTTGTTTTTGTTATTTTGTCATTTTGTTATTTTGCTATT
TTATGGATTAATTTTTGTTTATAAAATGGGAAATAATTTTAACATATTTAAATAAATGGA
GAAAAAATATAAAATAATTATAAAAAAAAGTTAATACACATTTTTTCCTGTTATAGACCT
TATATTTATTTATCCATATATATATATATATATATATATATATATATATACATACCAAGT
GAATTAAGAGGAAAGCTAATTTATTATTCAGAATAATATATGAACTATATATAATTTTTA
TTATTTTGGTGTATATTAATCTGTCTATATGCATACATGCAATAATTTATCGACTTATAT
ATCAAATAACATAAAATAGAAGTGTTTTAAATTATGGATATATGCTCAATATTCATTTTT
TTTAATAAGTTAGCTATATTTAAATTATACATTTTATATATGGTCTCTTTTTTTTTTAAA
TATTATTTAAGTGATCATGAAAATATAAATAATTTTTTTTTATTTAATATCCTTTTGCTT
GCATGTGGTAAATGGAAATTTGGATGTGTTTTGAARGTTCGGATATAGTTGTATGGACAT
ATAATATATTTTGTGAAAAATTGGTTTTATGTTTATACTTATGCCAATACTTTTTGAGTA
AAACAAAGCAAGTGCTTATAAATAATTAAAGCCAATTTTATAATATATATTTTTTTATTT
AATTTGAATTTAGTAGTATAATTTTTTATGGTAAGTGCTCAAAGAGAGTTGCTTATAAAG
TATGGTTTGTTTCTTTTTCGCCATTTTGAATTACACATTAAAAATATATAGATACATATA
TTATAATATGAAATCATTAATAATTTAGGGAAATTCTACAAATTTAAAAACGAATAAAAT
AATTGTTTTTCATCATGCCATAACACAATATTGATATATACATGTACAAACATTTTTTTT
ATTTGGAAAATATAAATTATATAAAAAAAAATGTATAGTATACAAAATGAGCATATTCAC
ACGGGGTGGACGTTCATTTTTTCATTTTTCCCCTGTTTTTTATGAGTATATGATAAAATT
TTATGAACATTTACACAAAATGAAAATGGATATATAGGAAAAATGGAGCGGTATTTCATT
TATCTTTGATTGTCATTTGGATATTATATTACCYTGGGTAGGCAATTAAAAATGTTAAAT
AACAATTTAAGGAAATTATATTTTATATATTAAAATTAACACTGTATTATATGATTCGCT
TATAAAAGCCACTCTTTCCCCATGCAAAGCTGTTTAATATCAATTTTAACAAATTACACA
CATGTTAATATATTTATATATATAATTTATATATTTATAATTTATATATTTATATTTTTA
TTATTTATATATTTATTATTTATTGTGTGTGTCAATTCGGGTAGGATATACCTCTTTTTT
ATTGTTTAAAGCGATTTGTATTCTAAAATATAAAGRATTTGAAAAAGAGAAAGATAGAAT
ATGATCCCATCATATATAGCCCTATAATTTTTATTTAGCAGCGAATTAATTTTTCTATTA
AGTTTATGTGTAATTAAAATAACGGAATATATATAATACAATAAAAAAGTGCATAAATTA
AAATTTTTTCAATTAAATTTTTTTTTTTAAGGGGTTATATAATATTAAATATATAAAATA
CGATTATATATTTTTGCTACAATTTTTTATATTAAGATATAAATAGTAAATAAATGGTAT
TATATGGCATGTAATATATAAATTTTTTCCAATTTTTATTTTATATACACTTTTCCTTTT
TTTGTCATAAAACTTAAACAATTTACACATTCATTTTAAAAATTGACTATTTGTTTCAAC
ATTTTTTGAGTTTCCGTTTTATAATAGTATTTTCATTTGTATATTGCTTATATATATAAA
TACACACCTAAATGTTACAAAGGATCAATGCATAAACCGGTGTGTCTGGTCGTCGCGATG
ACCCCCAAGAGGGGCATCGGCATCAACAACGGCCTCCCGTGGCCCCACTTGACCACAGAT
TTCAAACACTTTTCTCGTGTGACAAAAACGACGCCCGAAGAAGCCAGTCGCCTGAACGGG
TGGCTTCCCAGGAAATTTGCAAAGACGGGCGACTCTGGACTTCCCTCTCCATCAGTCGGC
AAGAGATTCAACGCCGTTGTCATGGGACGGAAAACCTGGGAAAGCATGCCTCGAAAGTTT
AGACCCCTCGTGGACAGATTGAACATCGTCGTTTCCTCTTCCCTCAAAGAAGAAGACATT
GCGGCGGAGAAGCCTCAAGCTGAAGGCCAGCAGCGCGTCCGAGTCTGTGCTTCACTCCCA
GCAGCTCTCAGCCTTCTGGAGGAAGAGTACAAGGATTCTGTCGACCAGATTTTTGTCGTG
GGAGGAGCGGGACTGTACGAGGCAGCGCTGTCTCTGGGCGTTGCCTCTCACCTGTACATC
ACGCGTGTAGCCCGCGAGTTTCCGTGCGACGTTTTCTTCCCTGCGTTCCCCGGAGATGAC
ATTCTTTCAAACAAATCAACTGCTGCGCAGGCTGCAGCTCCTGCCGAGTCTGTGTTCGTT
CCCTTTTGTCCGGAGCTCGGAAGAGAGAAGGACAATGAAGCGACGTATCGACCCATCTTC
ATTTCCAAGACCTTCTCAGACAACGGGGTTCCCTACGACTTTGTGGTTCTCGAGAAGAGA
AGGAAGACTGACGACGCAGCCACTGCGGAACCGAGCAACGCAATGAGCTCCTTGACGTCC
ACGAGGGAGACAACTCCCGTGCACGGGTTGCAGGCTCCTTCTTCGGCCGCAGCCATTGCC
CCGGTGTTGGCGTGGATGGACGAAGAAGACCGGAAAAAACGCGAGCAAAAGGAACTGATT
CGGGCCGTTCCGCATGTTCACTTTAGAGGCCATGAAGAGTTCCAGTACCTTGATCTCATT
GCCGACATTATTAACAATGGAAGGACAATGGATGACCGAACGGGCGTTGGTGTCATCTCC
AAATTCGGCTGCACTATGCGCTACTCGCTGGATCAGGCCTTTCCACTTCTCACCACAAAG
CGTGTGTTCTGGAAAGGGGTCCTCGAAGAGTTGCTGTGGTTCATTCGCGGCGACACGAAC
GCAAACCATCTTTCTGAGAAGGGCGTGAAGATCTGGGACAAGAATGTGACACGCGAGTTC
CTCGATTCGCGCAATCTCCCCCACCGAGAGGTCGGAGACATCGGCCCGGGCTACGGCTTC
CAGTGGAGACACTTCGGCGCGGCATACAAAGACATGCACACAGACTACACAGGGCAGGGC
GTCGACCAGCTGAAGAATGTGATCCAGATGCTGAGAACGAATCCAACAGATCGTCGCATG
CTCATGACTGCCTGGAATCCTGCAGCGCTGGACGAAATGGCGCTGCCGCCTTGTCACTTG
TTGTGCCAGTTCTACGTGAACGACCAGAAGGAGCTGTCGTGCATCATGTATCAGCGGTCG
TGCGATGTCGGCCTCGGCGTCCCCTTCAACATCGCTTCCTATTCGCTTTTGACGCTCATG
GTTGCACACGTCTGCAACCTAAAACCTAAGGAGTTCATTCACTTCATGGGGAACACGCAT
GTCTACACGAACCATGTCGAGGCTTTAAAAGAGCAGCTGCGGAGAGAACCGAGACCGTTC
CCCATTGTGAACATCCTCAACAAGGAACGCATCAAGGAAATCGACGATTTCACCGCCGAG
GATTTTGAGGTCGTGGGCTACGTCCCGCACGGACGAATCCAGATGGAGATGGCTGTCTAG
CGGAAATACAGAAGCTAGCTTTGATCCCGTTTTTCTTACTTATATATTTATACCAATTGA
TTGTATTTATAACTGTAAAAATGTGTATGTTGTGTGCATATTTTTTTTTGTGCATGCACA
TGCATGTAAATAGCTAAAATTATGAACATTTTATTTTTTGTTCAGAAAAAAAAAACTTTA
CACACATAAAATGGCTAGTATGAATAGCCATATTTTATATAAATTAAATCCTATGAATTT
ATGACCATATTAAAAATTTAGATATTTATGGAACATAATATGTTTGAAACAATAAGACAA
AATTATTATTATTATTATTATTTTTACTGTTATAATTATGTTGTCTCTTCAATGATTCAT
AAATAGTTGGACTTGATTTTTAAAATGTTTATAATATGATTAGCATAGTTAAATAAAAAA
AGTTGAAAAATTAAAAAAAAACATATAAACACAAATGATGTTTTTTCCTTCAATTTCGAT
ATCCCATAAGATTAGGATTGGAGCCAGTTGAGAAAATTGTAGCACAATCTATTCAACCTC
CTAAAGTCAACCGCCATACATACTCATTGGTGTCCCCAGTAGTAAAAGCACTTTTTAACA
AGATAGAAGAAGCCGTTCACAAGCCAGTTAGTGATAATATATGGGACTATGCAGGGGGTG
ATGATGAATATGAAGAAACTGAAGAAGACAACTTCGATAATGATTTCTTTAATTAAGACT
AATTTTATTTGAAAAAAATATACATTTAATATCAATGATAATAATAATAATAATGTAAAC
AATATTATTGTGTAACATTTTATAGTTTTTTGTATTGCAGAATTACAGAAACTATATTAT
ATTTTTAATATATATATACATACATATGTATATATAAGTGATGTTTTTTATAATATTATG
CTTTTTTACCTTATGATTTAAAAAATATGTTTTTTGTTTTTTCAATATTATGCTTATATT
TTAATTAATTTATTATATTTTTTATGTGGATTTGATTTCCTTATTTTAAACACATCTTTG
ATTTTAATAATAGAACATAGATATTAGATATTTTGAATTTTATTATAATTATATATTATT
TTTAATCCCATTTATTATTTTTTTTCTTTAAAAACAACATTAAGTGATAATCGTAACATT
TTTAATAACAAAATGCATTAAGACAATAATTTCCAGATAATTTGATAAAATAAAATCAAC
AAATAATGGCCAATTTAATTAATAACGGTATTATAGTTAAAATAAAGTTCATGCTAATTT
ATTAATTATATTTACAAAATAAGGAAATATGTGTATTAGACAGCAATTTCTATGTATTTT
TTATATACTATTAAAAGCTCAATTGCTCAAAAAATAAATAAGTATTTCTTCTATAAAAAA
TATCATCGAAAAAAATATATAAGATTACATTTAAAAAAACTTATTTACATTGGGTATTTT
AGGCAAAACGGGATATATATTCAGCTCAATGCACATCTAGTTATTACATTAAATATGAAA
AAAAAAAAACAATTTCATCTCAATCTAGCTAATACTTAACTAATATTTTTAAAATAAAGA
AAATACACCTTGCACATAAAAAAGAAGAAAAATTAAAAAAAAATTGTATATGTGCACACT
ATATTTATTAATTTTTTTTATCATAAAATATATAAATTGTGAAAATACACTAAAAATTGT
AGATTTTAGTAAAAATTACTTACGGATTAGATATGTGCAAAATACGAAAAATTAGCAAAT
GCAAAATAAATATAAAATTAATATCCATACTATATATATTTTAAGACAAATTTTTTTAAA
ACCAGCTTTTTTAAAATCGTAATTAGTGATATATAATTTAATGATATAAAAATATTTTTT
ATGTTTTACATATTTATGAGGCAATTATTTTCATTAGGGCTATCGTTTAATATATAAAAT
TTTAAAAATTGAAATAAACAAAATAAGTTATATAATCGTATAAGAAAATATATATTTATA
AAGATTAAACTATTTACATTATAATAATTAGTAAAAAAAAAATATATACTATTCGGATCC
ATGAGTAAAGGAGAAGAACTTTTCACTGGAGTTGTCCCAATTCTTGTTGAATTAGATGGT
GATGTTAATGGGCACAAATTTTCTGTCAGTGGAGAGGGTGAAGGTGATGCAACATACGGA
AAACTTACCCTTAAATTTATTTGCACTACTGGAAAACTACCTGTTCCATGGCCAACACTT
GTCACTACTTTCGGTTATGGTGTTCAATGCTTTGCGAGATACCCAGATCATATGAAACAG
CATGACTTTTTCAAGAGTGCCATGCCCGAAGGTTATGTACAGGAAAGAACTATATTTTTC
AAAGATGACGGGAACTACAAGACACGTGCTGAAGTCAAGTTTGAAGGTGATACCCTTGTT
AATAGAATCGAGTTAAAAGGTATTGATTTTAAAGAAGATGGAAACATTCTTGGACACAAA
TTGGAATACAACTATAACTCACACAATGTATACATCATGGCAGACAAACAAAAGAATGGA
ATCAAAGTTAACTTCAAAATTAGACACAACATTGAAGATGGAAGCGTTCAACTAGCAGAC
CATTATCAACAAAATACTCCAATTGGCGATGGCCCTGTCCTTTTACCAGACAACCATTAC
CTGTCCACACAATCTGCCCTTTCGAAAGATCCCAACGAAAAGAGAGACCACATGGTCCTT
CTTGAGTTTGTAACAGCTGCTGGGATTACACATGGCATGGATGAACTATACAAATAAATT
CTAGAAGATCCCGTTTTTCTTACTTATATATTTATACCAATTGATTGTATTTATAACTGT
AAAAATGTGTATGTTGTGTGCATATTTTTTTTTGTGCATGCACATGCATGTAAATAGCTA
AAATTATGAACATTTTATTTTTTGTTCAGAAAAAAAAAACTTTACACACATAAAATGGCT
AGTATGAATAGCCATATTTTATATAAATTAAATCCTATGAATTTATGACCATATTAAAAA
TTTAGATATTTATGGAACATAATATGTTTGAAACAATAAGACAAAATTATTATTATTATT
ATTATTTTTACTGTTATAATTATGTTGTCTCTTCAATGATTCATAAATAGTTGGACTTGA
TTTTTAAAATGTTTATAATATGATTAGCATAGTTAAATAAAAAAAGTTGAAAAATTAAAA
AAAAACATATAAACACAAATGATGTTTTTTCCTTCAATTTCGGGTACCGAGCTCGAATTC
AACCTGGTTGATCTTGCCAGTAGTCA
Restriction sites to linearize plasmid KspI
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
Other details transgene
Promoter
Gene Model of 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
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 1ATAGAgatatcCCATAAGATTAGGATTGGAGCC
Additional information primer 1L2465 (EcoRV); forward PB000248.02.0 putative promoter
Sequence Primer 2CGCggatccGAATAGTATATATTTTTTTTTTACTAATTATTATAATG
Additional information primer 2L2466 (BamHI); reverse PB000248.02.0 putative promoter
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 Not available
Gene productNot available
Gene product: Alternative namesmall subunit ribosomal rna gene (c-type unit)
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 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