Summary

RMgm-585
Malaria parasiteP. berghei
Genotype
DisruptedGene model (rodent): PBANKA_1300800; Gene model (P.falciparum): PF3D7_1475400; Gene product: cysteine repeat modular protein 4 (CRMP4)
Phenotype Oocyst; Sporozoite; Liver stage;
Last modified: 3 April 2011, 14:06
  *RMgm-585
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) : 21453484
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/ResearcherJ. Thompson; K.D. Augustijn
Name Group/DepartmentInstitute of Immunology and Infection Research
Name InstituteSchool of Biological Sciences, University of Edinburgh
CityEdinburgh
CountryUK
Name of the mutant parasite
RMgm numberRMgm-585
Principal name376cl1; 376cl2
Alternative nameΔpcrmp4
Standardized name
Is the mutant parasite cloned after genetic modificationYes
Phenotype
Asexual blood stageNot different from wild type
Gametocyte/GameteNot different from wild type
Fertilization and ookineteNot different from wild type
OocystNormal numbers of oocysts are produced. Inside oocysts morphologically normal sporozoites are formed. Sporozoites fail to egress from oocysts. No sporozoites are found in the salivary glands.
SporozoiteNormal numbers of oocysts are produced. Inside oocysts morphologically normal sporozoites are formed. Sporozoites fail to egress from oocysts. No sporozoites are found in the salivary glands.
Mechanically liberated sporozoites from oocysts show gliding motility and invade and infect hepatocytes but do not undergo further development.
Liver stageMechanically liberated sporozoites from oocysts show gliding motility and invade and infect hepatocytes in vitro but do not undergo further development.
Additional remarks phenotype

Mutant/mutation
The mutant lacks expression of CRMP4 (cysteine repeat modular protein 4)

Protein (function)
The Cysteine Repeat Modular Proteins (CRMP1–4) of Plasmodium, are encoded by a small gene family that is conserved in malaria and other Apicomplexan parasites. They are very large, predicted surface proteins with multipass transmembrane domains containing motifs that are conserved within families of cysteine-rich, predicted surface proteins in a range of unicellular eukaryotes, and a unique combination of protein-binding motifs, including a > 100 kDa cysteine-rich modular region, an epidermal growth factor-like domain and a Kringle domain.

Phenotype
The phenotype analyses indicate that CRMP4 is not essential for blood stage development and for the formation of oocysts. The phenotype analyses indicate that CRMP4 plays a role in egress of sporozoites from oocysts and also during development of the early liver stages.

Additional information
The phenotype of P. berghei mutants lacking expression of CRMP3 (RMgm-584) and CRMP4 is different from P. berghei mutants lacking expression of CRMP1 (RMgm-198) and CRMP2 (RMgm-199). Crmp1 and crmp2 gene knockout parasites  establish normal blood-stage infection and transmit to the mosquito to form oocysts that release large numbers of sporozoites into the mosquito haemocoel. These sporozoites are infectious to the mouse when injected intravenously but are unable to target and invade mosquito salivary glands and thus do not transmit via infected mosquito bite.

crmp4 was also disrupted in the GFP-reporter line P. berghei ANKA 507cl1 (RMgm-7) using the same method/plasmid as described here (line 659m1cl1). In addition, 'double knock' mutants have been generated lacking both CRMP3 and CRMP4 (line  656m2cl1, 656m2cl2). Details of these lines have not been published. CRMP4 has been tagged with GFP (line 478) and cmyc (line 1109) but parasites of these lines have not been analysed in detail.

Other mutants
RMgm-198: A mutant lacking expression of CRMP1
RMgm-199: A mutant lacking expression of CRMP2
RMgm-584: A mutant lacking expression of CRMP3


  Disrupted: Mutant parasite with a disrupted gene
Details of the target gene
Gene Model of Rodent Parasite PBANKA_1300800
Gene Model P. falciparum ortholog PF3D7_1475400
Gene productcysteine repeat modular protein 4
Gene product: Alternative nameCRMP4
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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AATTCATATGGTACAAACTATGGGATACGATGACCACACTATTAAAAATAAAAATTATAA
ATTTTATTATAAAGTTATGAATAAAATCAAATTTGTCTTAAAAAAAAAACATAAAGAAAG
TATATTGCAATACGCTAAAAATGATAGCTCTATAGAAAAAACACATGGTCAACCAAAAGA
ATACATACATTTTGAGGCAATTGATAATATTATCAAAAAATATAATGATAATTTAAAAAA
AACAGTAATAAGAAATATAGACCAAAACGAAGAAATAATAGACAACGAACAAAAAAATAT
AAAAGGAAAAGCACAATCTTCATTAAAAAAAAAAAAATTATCAGTTAATGTGTGGAATGT
AGCATATATATGGTATGATGAACAAAATGAAGATTTGTTTTTCCAAATGCCTTTTGAAGA
TAAGTTATCTTCAAGTTTTAATAAAGAACCCAATGAAAAAACATATAATATATTCGCTCG
TATACCAGATCGTATCAAAGATAAAGAAACAAAATATAATATCAAATATTTTGTACTAGC
TATAATACAAGTTATTGATAAATTCGTGGTTATATCAAAATCATGTTCTATATATGAAAA
TTGGTTTGATTTTGCAATGAGATTTGCATTTGTTTATATATCATGGATTAAAAGAATTAA
TAAAAAAGATATAATTATACCGAACACCATGGATCAATTACATGCTATAACAGATGAATA
TATATATATCCCTTTATTTTATAAATATGAGTATTTTACTCGCCCCAAAAAAGCATTTTA
TGAATCTCCAAAAGAAGAAACAAATTATGCTCCAAATAACTATGAAGAAGGAAAGTATCG
AACTGAAACATTGGAAAAAATGAATAGTTTCCAAAATACAGAAATCAAAATTAGTAGAAG
TTATGGATCATATAGCAAAATGAACAGTTTAAAGAATAGAAAAAAAGGGAAAGAAACAAA
TGATTATTTTAATGAGCAAAACAGTAGGGAAGACTCAAATGGATCCACTAGTTCTAGAGC
GGCCGCCACCGCGGTGGAGCTCCAATTCGCCCTATAGTGAGTCGTATTACGCGCGCTCAC
TGGCCGTCGTTTTACAACGTCGTGACTGGGAAAACCCTGGCGTTACCCAACTTAATCGCC
TTGCAGCACATCCCCCTTTCGCCAGCTGGCGTAATAGCGAAGAGGCCCGCACCGATCGCC
CTTCCCAACAGTTGCGCAGCCTGAATGGCGAATGGGACGCGCCCTGTAGCGGCGCATTAA
GCGCGGCGGGTGTGGTGGTTACGCGCAGCGTGACCGCTACACTTGCCAGCGCCCTAGCGC
CCGCTCCTTTCGCTTTCTTCCCTTCCTTTCTCGCCACGTTCGCCGGCTTTCCCCGTCAAG
CTCTAAATCGGGGGCTCCCTTTAGGGTTCCGATTTAGTGCTTTACGGCACCTCGACCCCA
AAAAACTTGATTAGGGTGATGGTTCACGTAGTGGGCCATCGCCCTGATAGACGGTTTTTC
GCCCTTTGACGTTGGAGTCCACGTTCTTTAATAGTGGACTCTTGTTCCAAACTGGAACAA
CACTCAACCCTATCTCGGTCTATTCTTTTGATTTATAAGGGATTTTGCCGATTTCGGCCT
ATTGGTTAAAAAATGAGCTGATTTAACAAAAATTTAACGCGAATTTTAACAAAATATTAA
CGCTTACAATTTAGGTGGCACTTTTCGGGGAAATGTGCGCGGAACCCCTATTTGTTTATT
TTTCTAAATACATTCAAATATGTATCCGCTCATGAGACAATAACCCTGATAAATGCTTCA
ATAATATTGAAAAAGGAAGAGTATGAGTATTCAACATTTCCGTGTCGCCCTTATTCCCTT
TTTTGCGGCATTTTGCCTTCCTGTTTTTGCTCACCCAGAAACGCTGGTGAAAGTAAAAGA
TGCTGAAGATCAGTTGGGTGCACGAGTGGGTTACATCGAACTGGATCTCAACAGCGGTAA
GATCCTTGAGAGTTTTCGCCCCGAAGAACGTTTTCCAATGATGAGCACTTTTAAAGTTCT
GCTATGTGGCGCGGTATTATCCCGTATTGACGCCGGGCAAGAGCAACTCGGTCGCCGCAT
ACACTATTCTCAGAATGACTTGGTTGAGTACTCACCAGTCACAGAAAAGCATCTTACGGA
TGGCATGACAGTAAGAGAATTATGCAGTGCTGCCATAACCATGAGTGATAACACTGCGGC
CAACTTACTTCTGACAACGATCGGAGGACCGAAGGAGCTAACCGCTTTTTTGCACAACAT
GGGGGATCATGTAACTCGCCTTGATCGTTGGGAACCGGAGCTGAATGAAGCCATACCAAA
CGACGAGCGTGACACCACGATGCCTGTAGCAATGGCAACAACGTTGCGCAAACTATTAAC
TGGCGAACTACTTACTCTAGCTTCCCGGCAACAATTAATAGACTGGATGGAGGCGGATAA
AGTTGCAGGACCACTTCTGCGCTCGGCCCTTCCGGCTGGCTGGTTTATTGCTGATAAATC
TGGAGCCGGTGAGCGTGGGTCTCGCGGTATCATTGCAGCACTGGGGCCAGATGGTAAGCC
CTCCCGTATCGTAGTTATCTACACGACGGGGAGTCAGGCAACTATGGATGAACGAAATAG
ACAGATCGCTGAGATAGGTGCCTCACTGATTAAGCATTGGTAACTGTCAGACCAAGTTTA
CTCATATATACTTTAGATTGATTTAAAACTTCATTTTTAATTTAAAAGGATCTAGGTGAA
GATCCTTTTTGATAATCTCATGACCAAAATCCCTTAACGTGAGTTTTCGTTCCACTGAGC
GTCAGACCCCGTAGAAAAGATCAAAGGATCTTCTTGAGATCCTTTTTTTCTGCGCGTAAT
CTGCTGCTTGCAAACAAAAAAACCACCGCTACCAGCGGTGGTTTGTTTGCCGGATCAAGA
GCTACCAACTCTTTTTCCGAAGGTAACTGGCTTCAGCAGAGCGCAGATACCAAATACTGT
CCTTCTAGTGTAGCCGTAGTTAGGCCACCACTTCAAGAACTCTGTAGCACCGCCTACATA
CCTCGCTCTGCTAATCCTGTTACCAGTGGCTGCTGCCAGTGGCGATAAGTCGTGTCTTAC
CGGGTTGGACTCAAGACGATAGTTACCGGATAAGGCGCAGCGGTCGGGCTGAACGGGGGG
TTCGTGCACACAGCCCAGCTTGGAGCGAACGACCTACACCGAACTGAGATACCTACAGCG
TGAGCTATGAGAAAGCGCCACGCTTCCCGAAGGGAGAAAGGCGGACAGGTATCCGGTAAG
CGGCAGGGTCGGAACAGGAGAGCGCACGAGGGAGCTTCCAGGGGGAAACGCCTGGTATCT
TTATAGTCCTGTCGGGTTTCGCCACCTCTGACTTGAGCGTCGATTTTTGTGATGCTCGTC
AGGGGGGCGGAGCCTATGGAAAAACGCCAGCAACGCGGCCTTTTTACGGTTCCTGGCCTT
TTGCTGGCCTTTTGCTCACATGTTCTTTCCTGCGTTATCCCCTGATTCTGTGGATAACCG
TATTACCGCCTTTGAGTGAGCTGATACCGCTCGCCGCAGCCGAACGACCGAGCGCAGCGA
GTCAGTGAGCGAGGAAGCGGAAGAGCGCCCAATACGCAAACCGCCTCTCCCCGCGCGTTG
GCCGATTCATTAATGCAGCTGGCACGACAGGTTTCCCGACTGGAAAGCGGGCAGTGAGCG
CAACGCAATTAATGTGAGTTAGCTCACTCATTAGGCACCCCAGGCTTTACACTTTATGCT
TCCGGCTCGTATGTTGTGTGGAATTGTGAGCGGATAACAATTTCACACAGGAAACAGCTA
TGACCATGATTACGCCAAGCTCGAAATTAACCCTCACTAAAGGGAACAAAAGCTGGTACC
TTGGCATCTTCTATTTTCGATTGTGTTTGTAAACGTGGTTATGGAACATTAAATGAGAGA
AGAAATGGTATTAAAAAATCCTTAACATGTAAATTATGTCCCAAAAATTTCTTTAAACAC
AATACAACCGATGAATACTGTTTACCTTGCCCCTATAAAACTTATACATTATCAGAAGGA
TCAAATTCGATACGCAGTTGTTTGCCTAGAGAAGGGCATTTTATACTAATGCTTAGAAAT
ATAAATGTAGAATATTTAAGGTATAAATTTGATTCGAATATGGAAAACACAATTTATAAT
TACTTTCAAAATAATAGCAAAAGTTTTGATGATGATGTGTATGACTTATTTTTAAATAAC
AATTTTGAGCAGGTATTTCAAAATTTGAATAATAACACCAAAACAAATAACAGCATTAAT
AAAAAAAATGGAAATAAAGAAGAAAATTTATATCCCAAAGAAGATAAAAATCAAAATGCC
ATCAACAATAACCCCAACAATCAAAATAATAACAATAATAAATACAATTTAAAGGATTTG
AAAAAGCTCGGGTTTCTTGAACCAAACTTAATATATTATTTATATTCAATAATGTTAAGA
AATATAGAAGATAATTATAACTGGAAAAATGACAATAATTTAAATGTTATTTGCCATATT
AATGTTCACTCGAAAAATAACCCTAATTTTACAGTTACATATACACCAAATTTGAGCTCG
TGCATATCCAGTTGTAAAAGTAATATATATTGTACTGGTGTCGAATTTTCTCGGTATCGA
GTTGAATACGAACAAATTTTTTTAAAAAATAATCAAAAGATTATTGTTGGATATTTTAAA
TGCAAAAAATTTTATTATAAAGATATATATTATTATTCTTCATCTAATTTAGAACAATTT
TTGAATGAAAACGTTCAGAATGGTGTACCAATAAAGAAGCTTGCATGCCTGCAGGTCAAC
AATAAATAATAAATAAATATTGTGGAAATAAAATAACATATAATTATTTTTAATACATTG
ATTTCCCTTTTATTTTTTTAAATTTCATTGATATAAAAATATATAATAATAACATATATG
ATTTCAAATTAATCTTTTCAAAAATGGTGTTTATTTTTGTATGTTTGTGTATGAATTAAT
CACATAACACATCTATTAAATTGAGTTGGTAATATAGACACAAATAAATATATATATTTT
TATAGCTTAAAAGTGTGTTATGAATATTTTAAGCATATTTTCTTTTTCTTTGGATTGTGT
AAAATGAACTCATATAATGCGTTTTTTTGTTTTTGTTATTTTGTCATTTTGTTATTTTGC
TATTTTATGGATTAATTTTTGTTTATAAAATGGGAAATAATTTTAACATATTTAAATAAA
TGGAGAAAAAATATAAAATAATTATAAAAAAAAGTTAATACACATTTTTTCCTGTTATAG
ACCTTATATTTATTTATCCATATATATATATATATATATATATATATATATATACATACC
AAGTGAATTAAGAGGAAAGCTAATTTATTATTCAGAATAATATATGAACTATATATAATT
TTTATTATTTTGGTGTATATTAATCTGTCTATATGCATACATGCAATAATTTATCGACTT
ATATATCAAATAACATAAAATAGAAGTGTTTTAAATTATGGATATATGCTCAATATTCAT
TTTTTTTAATAAGTTAGCTATATTTAAATTATACATTTTATATATGGTCTCTTTTTTTTT
TAAATATTATTTAAGTGATCATGAAAATATAAATAATTTTTTTTTATTTAATATCCTTTT
GCTTGCATGTGGTAAATGGAAATTTGGATGTGTTTTGAARGTTCGGATATAGTTGTATGG
ACATATAATATATTTTGTGAAAAATTGGTTTTATGTTTATACTTATGCCAATACTTTTTG
AGTAAAACAAAGCAAGTGCTTATAAATAATTAAAGCCAATTTTATAATATATATTTTTTT
ATTTAATTTGAATTTAGTAGTATAATTTTTTATGGTAAGTGCTCAAAGAGAGTTGCTTAT
AAAGTATGGTTTGTTTCTTTTTCGCCATTTTGAATTACACATTAAAAATATATAGATACA
TATATTATAATATGAAATCATTAATAATTTAGGGAAATTCTACAAATTTAAAAACGAATA
AAATAATTGTTTTTCATCATGCCATAACACAATATTGATATATACATGTACAAACATTTT
TTTTATTTGGAAAATATAAATTATATAAAAAAAAATGTATAGTATACAAAATGAGCATAT
TCACACGGGGTGGACGTTCATTTTTTCATTTTTCCCCTGTTTTTTATGAGTATATGATAA
AATTTTATGAACATTTACACAAAATGAAAATGGATATATAGGAAAAATGGAGCGGTATTT
CATTTATCTTTGATTGTCATTTGGATATTATATTACCYTGGGTAGGCAATTAAAAATGTT
AAATAACAATTTAAGGAAATTATATTTTATATATTAAAATTAACACTGTATTATATGATT
CGCTTATAAAAGCCACTCTTTCCCCATGCAAAGCTGTTTAATATCAATTTTAACAAATTA
CACACATGTTAATATATTTATATATATAATTTATATATTTATAATTTATATATTTATATT
TTTATTATTTATATATTTATTATTTATTGTGTGTGTCAATTCGGGTAGGATATACCTCTT
TTTTATTGTTTAAAGCGATTTGTATTCTAAAATATAAAGRATTTGAAAAAGAGAAAGATA
GAATATGATCCCATCATATATAGCCCTATAATTTTTATTTAGCAGCGAATTAATTTTTCT
ATTAAGTTTATGTGTAATTAAAATAACGGAATATATATAATACAATAAAAAAGTGCATAA
ATTAAAATTTTTTCAATTAAATTTTTTTTTTTAAGGGGTTATATAATATTAAATATATAA
AATACGATTATATATTTTTGCTACAATTTTTTATATTAAGATATAAATAGTAAATAAATG
GTATTATATGGCATGTAATATATAAATTTTTTCCAATTTTTATTTTATATACACTTTTCC
TTTTTTTGTCATAAAACTTAAACAATTTACACATTCATTTTAAAAATTGACTATTTGTTT
CAACATTTTTTGAGTTTCCGTTTTATAATAGTATTTTCATTTGTATATTGCTTATATATA
TAAATACACACCTAAATGTTACAAAGGATCAATGCATAAACCGGTGTGTCTGGTCGTCGC
GATGACCCCCAAGAGGGGCATCGGCATCAACAACGGCCTCCCGTGGCCCCACTTGACCAC
AGATTTCAAACACTTTTCTCGTGTGACAAAAACGACGCCCGAAGAAGCCAGTCGCCTGAA
CGGGTGGCTTCCCAGGAAATTTGCAAAGACGGGCGACTCTGGACTTCCCTCTCCATCAGT
CGGCAAGAGATTCAACGCCGTTGTCATGGGACGGAAAACCTGGGAAAGCATGCCTCGAAA
GTTTAGACCCCTCGTGGACAGATTGAACATCGTCGTTTCCTCTTCCCTCAAAGAAGAAGA
CATTGCGGCGGAGAAGCCTCAAGCTGAAGGCCAGCAGCGCGTCCGAGTCTGTGCTTCACT
CCCAGCAGCTCTCAGCCTTCTGGAGGAAGAGTACAAGGATTCTGTCGACCAGATTTTTGT
CGTGGGAGGAGCGGGACTGTACGAGGCAGCGCTGTCTCTGGGCGTTGCCTCTCACCTGTA
CATCACGCGTGTAGCCCGCGAGTTTCCGTGCGACGTTTTCTTCCCTGCGTTCCCCGGAGA
TGACATTCTTTCAAACAAATCAACTGCTGCGCAGGCTGCAGCTCCTGCCGAGTCTGTGTT
CGTTCCCTTTTGTCCGGAGCTCGGAAGAGAGAAGGACAATGAAGCGACGTATCGACCCAT
CTTCATTTCCAAGACCTTCTCAGACAACGGGGTTCCCTACGACTTTGTGGTTCTCGAGAA
GAGAAGGAAGACTGACGACGCAGCCACTGCGGAACCGAGCAACGCAATGAGCTCCTTGAC
GTCCACGAGGGAGACAACTCCCGTGCACGGGTTGCAGGCTCCTTCTTCGGCCGCAGCCAT
TGCCCCGGTGTTGGCGTGGATGGACGAAGAAGACCGGAAAAAACGCGAGCAAAAGGAACT
GATTCGGGCCGTTCCGCATGTTCACTTTAGAGGCCATGAAGAGTTCCAGTACCTTGATCT
CATTGCCGACATTATTAACAATGGAAGGACAATGGATGACCGAACGGGCGTTGGTGTCAT
CTCCAAATTCGGCTGCACTATGCGCTACTCGCTGGATCAGGCCTTTCCACTTCTCACCAC
AAAGCGTGTGTTCTGGAAAGGGGTCCTCGAAGAGTTGCTGTGGTTCATTCGCGGCGACAC
GAACGCAAACCATCTTTCTGAGAAGGGCGTGAAGATCTGGGACAAGAATGTGACACGCGA
GTTCCTCGATTCGCGCAATCTCCCCCACCGAGAGGTCGGAGACATCGGCCCGGGCTACGG
CTTCCAGTGGAGACACTTCGGCGCGGCATACAAAGACATGCACACAGACTACACAGGGCA
GGGCGTCGACCAGCTGAAGAATGTGATCCAGATGCTGAGAACGAATCCAACAGATCGTCG
CATGCTCATGACTGCCTGGAATCCTGCAGCGCTGGACGAAATGGCGCTGCCGCCTTGTCA
CTTGTTGTGCCAGTTCTACGTGAACGACCAGAAGGAGCTGTCGTGCATCATGTATCAGCG
GTCGTGCGATGTCGGCCTCGGCGTCCCCTTCAACATCGCTTCCTATTCGCTTTTGACGCT
CATGGTTGCACACGTCTGCAACCTAAAACCTAAGGAGTTCATTCACTTCATGGGGAACAC
GCATGTCTACACGAACCATGTCGAGGCTTTAAAAGAGCAGCTGCGGAGAGAACCGAGACC
GTTCCCCATTGTGAACATCCTCAACAAGGAACGCATCAAGGAAATCGACGATTTCACCGC
CGAGGATTTTGAGGTCGTGGGCTACGTCCCGCACGGACGAATCCAGATGGAGATGGCTGT
CTAGCGGAAATACAGAAGCTAGCTTTGATCCCGTTTTTCTTACTTATATATTTATACCAA
TTGATTGTATTTATAACTGTAAAAATGTGTATGTTGTGTGCATATTTTTTTTTGTGCATG
CACATGCATGTAAATAGCTAAAATTATGAACATTTTATTTTTTGTTCAGAAAAAAAAAAC
TTTACACACATAAAATGGCTAGTATGAATAGCCATATTTTATATAAATTAAATCCTATGA
ATTTATGACCATATTAAAAATTTAGATATTTATGGAACATAATATGTTTGAAACAATAAG
ACAAAATTATTATTATTATTATTATTTTTACTGTTATAATTATGTTGTCTCTTCAATGAT
TCATAAATAGTTGGACTTGATTTTTAAAATGTTTATAATATGATTAGCATAGTTAAATAA
AAAAAGTTGAAAAATTAAAAAAAAACATATAAACACAAATGATGTTTTTTCCTTCAATTT
CGATATCG
Restriction sites to linearize plasmid KpnI, BamHI
Partial or complete disruption of the genePartial
Additional remarks partial/complete disruption CRM-4 cds: 16360bp
Disrupted region: 10969 - 14594
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 1GTAAATGGTACCTTGGCATCTTCTATTTTCGATTGTG
Additional information primer 1L1457 (KpnI); 5'target forward
Sequence Primer 2ATCTTCAAGCTTCTTTATTGGTACACCATTCTGAACG
Additional information primer 2L1458 (HindIII); 5'target reverse
Sequence Primer 3TCAAAGAATTCATATGGTACAAACTATGGGATACGATGACCAC
Additional information primer 3L1459 (EcoRI/NdeI); 3'target forward
Sequence Primer 4TTACCGGATCCATTTGAGTCTTCCCTACTGTTTTGCTC
Additional information primer 4L1461 (BamHI); 3'target reverse
Sequence Primer 5
Additional information primer 5
Sequence Primer 6
Additional information primer 6