RMgmDB - Rodent Malaria genetically modified Parasites

Summary

RMgm-628

Malaria parasite	P. berghei
Genotype
Tagged	Gene model (rodent): PBANKA_0932000; Gene model (P.falciparum): PF3D7_1116000; Gene product: rhoptry neck protein 4 (RON4) Name tag: mCherry
Phenotype	Asexual bloodstage; Sporozoite;

Last modified: 30 September 2020, 12:59

*RMgm-628

Successful modification	The parasite was generated by the genetic modification
The mutant contains the following genetic modification(s)	Gene tagging
Reference (PubMed-PMID number)	Reference 1 (PMID number) : 32979009
MR4 number
top of page
Parent parasite used to introduce the genetic modification
Rodent Malaria Parasite	P. berghei
Parent strain/line	P. berghei ANKA
Name parent line/clone	P. berghei ANKA cl15cy1
Other information parent line	A reference wild type clone from the ANKA strain of P. berghei (PubMed: PMID: 17406255).
top of page
The mutant parasite was generated by
Name PI/Researcher	C.J. Janse; O. Klop; B.M. Franke-Fayard
Name Group/Department	Leiden Malaria Research Group
Name Institute	Leiden University Medical Center
City	Leiden
Country	The Netherlands
top of page
Name of the mutant parasite
RMgm number	RMgm-628
Principal name	1964
Alternative name	RON4::mCherry
Standardized name
Is the mutant parasite cloned after genetic modification	No
top of page
Phenotype
Asexual blood stage	Schizonts express mCherry-tagged RON4 as shown by fluorescence microscopy. Merozoites of mature schizonts show a 'rhoptry-like' fluorescence pattern (see also 'Additional remarks phenotype').
Gametocyte/Gamete	Not tested
Fertilization and ookinete	Not tested
Oocyst	Not tested
Sporozoite	RON4::mCherry expression in sporozoites
Liver stage	Not tested
Additional remarks phenotype	Mutant/mutation The mutant expresses an mCherry-tagged version of RON4 (rhoptry neck protein 4) Protein (function) RON4 forms a complex with different other proteins (e.g. AMA1, RON2, RON5) during invasion of the merozoite at the tight junction, an aperture in the host cell through which the parasite pulls itself before settling into a newly formed parasitophorous vacuole. Phenotype Both the genotype and phenotype of this mutant has not been analysed in detail. The mutant parasites have not been cloned after transfection and drug-selection. Schizonts express mCherry-tagged RON4 as shown by fluorescence microscopy. Merozoites of mature schizonts show a 'rhoptry-like' fluorescence pattern. The mutant can be obtained from the Leiden Malaria Research Group. The mutant has been (described and) used to analyse sporozoite migration in hepatocytes (Published in PMID: 32979009) Additional information Figure: Southern analysis of PFG-separated schizonts of transfected parasites after drug-selection Other mutants RMgm-214: Unsuccessful attempts to disrupt the ron2 gene (PBANKA_131570) RMgm-213: A mutant expressing a cmyc-tagged RON2 protein (PBANKA_131570)

Tagged: Mutant parasite with a tagged gene

top of page

Details of the target gene

Gene Model of Rodent Parasite PBANKA_0932000

Gene Model P. falciparum ortholog PF3D7_1116000

Gene product rhoptry neck protein 4

Gene product: Alternative name RON4

top of page

Details of the genetic modification

Name of the tag mCherry

Details of tagging C-terminal

Additional remarks: tagging

Commercial source of tag-antibodies

Type of plasmid/construct (Linear) plasmid single cross-over

PlasmoGEM (Sanger) construct/vector used No

Modified PlasmoGEM construct/vector used No

Plasmid/construct map

Plasmid/construct sequence

CTAAATTGTAAGCGTTAATATTTTGTTAAAATTCGCGTTAAATTTTTGTTAAATCAGCTC

ATTTTTTAACCAATAGGCCGAAATCGGCAAAATCCCTTATAAATCAAAAGAATAGACCGA

GATAGGGTTGAGTGTTGTTCCAGTTTGGAACAAGAGTCCACTATTAAAGAACGTGGACTC

CAACGTCAAAGGGCGAAAAACCGTCTATCAGGGCGATGGCCCACTACGTGAACCATCACC

CTAATCAAGTTTTTTGGGGTCGAGGTGCCGTAAAGCACTAAATCGGAACCCTAAAGGGAG

CCCCCGATTTAGAGCTTGACGGGGAAAGCCGGCGAACGTGGCGAGAAAGGAAGGGAAGAA

AGCGAAAGGAGCGGGCGCTAGGGCGCTGGCAAGTGTAGCGGTCACGCTGCGCGTAACCAC

CACACCCGCCGCGCTTAATGCGCCGCTACAGGGCGCGTCCCATTCGCCATTCAGGCTGCG

CAACTGTTGGGAAGGGCGATCGGTGCGGGCCTCTTCGCTATTACGCCAGCTGGCGAAAGG

GGGATGTGCTGCAAGGCGATTAAGTTGGGTAACGCCAGGGTTTTCCCAGTCACGACGTTG

TAAAACGACGGCCAGTGAGCGCGCGTAATACGACTCACTATAGGGCGAATTGGAGCTCCA

CCGCGGTGGCGGCCGCTCTAGAACTAGTGCATCCATATTAATGCAACAATGGAAATATTA

TCAAAATGCTTTAGGGTTTGAAGATAATAATGATCATGCATTTTTAGGTTTAATCGATGA

ATTATTAACAATAGACAAAAGATATAGTAAAAATGAAGAACATAGTAAATCTTTAGAACA

AATGAAAAAATCAAAAGTATTTAATTATTGTACTAAACTTATGAGAACAAGTGGAAACAT

TTCAACAATACCATTTAATCATGAAAACAATAAAACTCCAAGTTCATCCATTATTGGTTC

ATTAGGAAATTTAGTTAAGGCACATATAAGTGGATATTATGCAGCTACTGCTCAAAGAAT

TAACTCTTATTTTAATTATTCTAACAATAAAGATCAAAAGAAAGCACTATACAAAGTCAT

ATCAGTTTGTACTCTCTTAAATTTAACAGATATTTTATTTAATTGCACAGATCCACAATC

AACTAATTCAGCAAATTTGAACATTTTGAAGCTAAATGTATTAAATACTAAAGGAAAAGA

AATTCTATATCGCCTTTATAATTTAAGCTACTTATTAGAGAGCGAAAACGAAGCAATTAA

AGAAGTGTGTAATCCTAAAAATGCCTCAGTTGGTAATGCTAAAATGCGATGAATTATTTC

TTTGTATATTTTTTTGTGTGGATATGTGCTTTTTTTGTAGGTGTTATACGGGATTTCAAA

TATTGCAAAATATGCATATATTTCATATGCAATATTTTCGAATTTCATTTTATCATTTGA

TATGAACTTATATACATATTTGCGCTTGTTTGATTTTCATAAGAATATATGTATTTATAA

TTTCACTTTTATTTAATTATTTTTTACAGATAAATCGTTACAAAAATTGTTAGAATTACT

TTCATCAAACTCACACAACGTATTAGCACATGAAATGGAAAAGAGAGCTCTTGATACTGA

TTATATTAACGAGGAAATAAACAATATAAGTGATTTTGATCATAATGAAAAAGACAATGG

AGAAGATGATGTTGAGAAAGCCATTTTTGATGATTTAGGATCCGGTGGAGGTGGAAGATC

CATGGTGAGCAAGGGCGAGGAGGATAACATGGCCATCATCAAGGAGTTCATGCGCTTCAA

GGTGCACATGGAGGGCTCCGTGAACGGCCACGAGTTCGAGATCGAGGGCGAGGGCGAGGG

CCGCCCCTACGAGGGCACCCAGACCGCCAAGCTGAAGGTGACCAAGGGTGGCCCCCTGCC

CTTCGCCTGGGACATCCTGTCCCCTCAGTTCATGTACGGCTCCAAGGCCTACGTGAAGCA

CCCCGCCGACATCCCCGACTACTTGAAGCTGTCCTTCCCCGAGGGCTTCAAGTGGGAGCG

CGTGATGAACTTCGAGGACGGCGGCGTGGTGACCGTGACCCAGGACTCCTCCCTGCAGGA

CGGCGAGTTCATCTACAAGGTGAAGCTGCGCGGCACCAACTTCCCCTCCGACGGCCCCGT

AATGCAGAAGAAGACCATGGGCTGGGAGGCCTCCTCCGAGCGGATGTACCCCGAGGACGG

CGCCCTGAAGGGCGAGATCAAGCAGAGGCTGAAGCTGAAGGACGGCGGCCACTACGACGC

TGAGGTCAAGACCACCTACAAGGCCAAGAAGCCCGTGCAGCTGCCCGGCGCCTACAACGT

CAACATCAAGTTGGACATCACCTCCCACAACGAGGACTACACCATCGTGGAACAGTACGA

ACGCGCCGAGGGCCGCCACTCCACCGGCGGCATGGACGAGCTGTACAAGTAATCTAGAAT

TATTAATATATATGAATATATATACATCGTTGTATGCGTAATATACGTGTAACTGTTTTT

TTTAATTTTTATATTAACTTTTCCTTATACATTTTAATGCCATTAACTAATAATATTTTA

CTTGAAAAAATTCTAAATTTTTATAACTCTAATATAAAATCTGTGAATAATAAAAAATTA

GCATAAAGTACGAATTAAACATCGAATATTTGTTTTAAAGGGGCACACATTATGTGCATA

TAATTTTATTCTTTGAATTTAATTTTTTGCTTATAATTAGATTAAATTAATATAATTTTT

TTATTGAAAATAAAAACTTATTTTAAAAAATTAACAAATGGAAGTTAATGAAAAATACGA

AAAAAGTAAAGGGTTAATTCTTATATGGTCGGTACCGAGCTCGAATTCGATATCGAAATT

GAAGGAAAAAACATCATTTGTGTTTATATGTTTTTTTTTAATTTTTCAACTTTTTTTATT

TAACTATGCTAATCATATTATAAACATTTTAAAAATCAAGTCCAACTATTTATGAATCAT

TGAAGAGACAACATAATTATAACAGTAAAAATAATAATAATAATAATAATTTTGTCTTAT

TGTTTCAAACATATTATGTTCCATAAATATCTAAATTTTTAATATGGTCATAAATTCATA

GGATTTAATTTATATAAAATATGGCTATTCATACTAGCCATTTTATGTGTGTAAAGTTTT

TTTTTTCTGAACAAAAAATAAAATGTTCATAATTTTAGCTATTTACATGCATGTGCATGC

ACAAAAAAAAATATGCACACAACATACACATTTTTACAGTTATAAATACAATCAATTGGT

ATAAATATATAAGTAAGAAAAACGGGATCAAAGCTAGCTTCTGTATTTCCGCTAGACAGC

CATCTCCATCTGGATTCGTCCGTGCGGGACGTAGCCCACGACCTCAAAATCCTCGGCGGT

GAAATCGTCGATTTCCTTGATGCGTTCCTTGTTGAGGATGTTCACAATGGGGAACGGTCT

CGGTTCTCTCCGCAGCTGCTCTTTTAAAGCCTCGACATGGTTCGTGTAGACATGCGTGTT

CCCCATGAAGTGAATGAACTCCTTAGGTTTTAGGTTGCAGACGTGTGCAACCATGAGCGT

CAAAAGCGAATAGGAAGCGATGTTGAAGGGGACGCCGAGGCCGACATCGCACGACCGCTG

ATACATGATGCACGACAGCTCCTTCTGGTCGTTCACGTAGAACTGGCACAACAAGTGACA

AGGCGGCAGCGCCATTTCGTCCAGCGCTGCAGGATTCCAGGCAGTCATGAGCATGCGACG

ATCTGTTGGATTCGTTCTCAGCATCTGGATCACATTCTTCAGCTGGTCGACGCCCTGCCC

TGTGTAGTCTGTGTGCATGTCTTTGTATGCCGCGCCGAAGTGTCTCCACTGGAAGCCGTA

GCCCGGGCCGATGTCTCCGACCTCTCGGTGGGGGAGATTGCGCGAATCGAGGAACTCGCG

TGTCACATTCTTGTCCCAGATCTTCACGCCCTTCTCAGAAAGATGGTTTGCGTTCGTGTC

GCCGCGAATGAACCACAGCAACTCTTCGAGGACCCCTTTCCAGAACACACGCTTTGTGGT

GAGAAGTGGAAAGGCCTGATCCAGCGAGTAGCGCATAGTGCAGCCGAATTTGGAGATGAC

ACCAACGCCCGTTCGGTCATCCATTGTCCTTCCATTGTTAATAATGTCGGCAATGAGATC

AAGGTACTGGAACTCTTCATGGCCTCTAAAGTGAACATGCGGAACGGCCCGAATCAGTTC

CTTTTGCTCGCGTTTTTTCCGGTCTTCTTCGTCCATCCACGCCAACACCGGGGCAATGGC

TGCGGCCGAAGAAGGAGCCTGCAACCCGTGCACGGGAGTTGTCTCCCTCGTGGACGTCAA

GGAGCTCATTGCGTTGCTCGGTTCCGCAGTGGCTGCGTCGTCAGTCTTCCTTCTCTTCTC

GAGAACCACAAAGTCGTAGGGAACCCCGTTGTCTGAGAAGGTCTTGGAAATGAAGATGGG

TCGATACGTCGCTTCATTGTCCTTCTCTCTTCCGAGCTCCGGACAAAAGGGAACGAACAC

AGACTCGGCAGGAGCTGCAGCCTGCGCAGCAGTTGATTTGTTTGAAAGAATGTCATCTCC

GGGGAACGCAGGGAAGAAAACGTCGCACGGAAACTCGCGGGCTACACGCGTGATGTACAG

GTGAGAGGCAACGCCCAGAGACAGCGCTGCCTCGTACAGTCCCGCTCCTCCCACGACAAA

AATCTGGTCGACAGAATCCTTGTACTCTTCCTCCAGAAGGCTGAGAGCTGCTGGGAGTGA

AGCACAGACTCGGACGCGCTGCTGGCCTTCAGCTTGAGGCTTCTCCGCCGCAATGTCTTC

TTCTTTGAGGGAAGAGGAAACGACGATGTTCAATCTGTCCACGAGGGGTCTAAACTTTCG

AGGCATGCTTTCCCAGGTTTTCCGTCCCATGACAACGGCGTTGAATCTCTTGCCGACTGA

TGGAGAGGGAAGTCCAGAGTCGCCCGTCTTTGCAAATTTCCTGGGAAGCCACCCGTTCAG

GCGACTGGCTTCTTCGGGCGTCGTTTTTGTCACACGAGAAAAGTGTTTGAAATCTGTGGT

CAAGTGGGGCCACGGGAGGCCGTTGTTGATGCCGATGCCCCTCTTGGGGGTCATCGCGAC

GACCAGACACACCGGTTTATGCATTGATCCTTTGTAACATTTAGGTGTGTATTTATATAT

ATAAGCAATATACAAATGAAAATACTATTATAAAACGGAAACTCAAAAAATGTTGAAACA

AATAGTCAATTTTTAAAATGAATGTGTAAATTGTTTAAGTTTTATGACAAAAAAAGGAAA

AGTGTATATAAAATAAAAATTGGAAAAAATTTATATATTACATGCCATATAATACCATTT

ATTTACTATTTATATCTTAATATAAAAAATTGTAGCAAAAATATATAATCGTATTTTATA

TATTTAATATTATATAACCCCTTAAAAAAAAAAATTTAATTGAAAAAATTTTAATTTATG

CACTTTTTTATTGTATTATATATATTCCGTTATTTTAATTACACATAAACTTAATAGAAA

AATTAATTCGCTGCTAAATAAAAATTATAGGGCTATATATGATGGGATCATATTCTATCT

TTCTCTTTTTCAAATYCTTTATATTTTAGAATACAAATCGCTTTAAACAATAAAAAAGAG

GTATATCCTACCCGAATTGACACACACAATAAATAATAAATATATAAATAATAAAAATAT

AAATATATAAATTATAAATATATAAATTATATATATAAATATATTAACATGTGTGTAATT

TGTTAAAATTGATATTAAACAGCTTTGCATGGGGAAAGAGTGGCTTTTATAAGCGAATCA

TATAATACAGTGTTAATTTTAATATATAAAATATAATTTCCTTAAATTGTTATTTAACAT

TTTTAATTGCCTACCCARGGTAATATAATATCCAAATGACAATCAAAGATAAATGAAATA

CCGCTCCATTTTTCCTATATATCCATTTTCATTTTGTGTAAATGTTCATAAAATTTTATC

ATATACTCATAAAAAACAGGGGAAAAATGAAAAAATGAACGTCCACCCCGTGTGAATATG

CTCATTTTGTATACTATACATTTTTTTTTATATAATTTATATTTTCCAAATAAAAAAAAT

GTTTGTACATGTATATATCAATATTGTGTTATGGCATGATGAAAAACAATTATTTTATTC

GTTTTTAAATTTGTAGAATTTCCCTAAATTATTAATGATTTCATATTATAATATATGTAT

CTATATATTTTTAATGTGTAATTCAAAATGGCGAAAAAGAAACAAACCATACTTTATAAG

CAACTCTCTTTGAGCACTTACCATAAAAAATTATACTACTAAATTCAAATTAAATAAAAA

AATATATATTATAAAATTGGCTTTAATTATTTATAAGCACTTGCTTTGTTTTACTCAAAA

AGTATTGGCATAAGTATAAACATAAAACCAATTTTTCACAAAATATATTATATGTCCATA

CAACTATATCCGAACYTTCAAAACACATCCAAATTTCCATTTACCACATGCAAGCAAAAG

GATATTAAATAAAAAAAAATTATTTATATTTTCATGATCACTTAAATAATATTTAAAAAA

AAAAGAGACCATATATAAAATGTATAATTTAAATATAGCTAACTTATTAAAAAAAATGAA

TATTGAGCATATATCCATAATTTAAAACACTTCTATTTTATGTTATTTGATATATAAGTC

GATAAATTATTGCATGTATGCATATAGACAGATTAATATACACCAAAATAATAAAAATTA

TATATAGTTCATATATTATTCTGAATAATAAATTAGCTTTCCTCTTAATTCACTTGGTAT

GTATATATATATATATATATATATATATATATATATGGATAAATAAATATAAGGTCTATA

ACAGGAAAAAATGTGTATTAACTTTTTTTTATAATTATTTTATATTTTTTCTCCATTTAT

TTAAATATGTTAAAATTATTTCCCATTTTATAAACAAAAATTAATCCATAAAATAGCAAA

ATAACAAAATGACAAAATAACAAAAACAAAAAAACGCATTATATGAGTTCATTTTACACA

ATCCAAAGAAAAAGAAAATATGCTTAAAATATTCATAACACACTTTTAAGCTATAAAAAT

ATATATATTTATTTGTGTCTATATTACCAACTCAATTTAATAGATGTGTTATGTGATTAA

TTCATACACAAACATACAAAAATAAACACCATTTTTGAAAAGATTAATTTGAAATCATAT

ATGTTATTATTATATATTTTTATATCAATGAAATTTAAAAAAATAAAAGGGAAATCAATG

TATTAAAAATAATTATATGTTATTTTATTTCCACAATATTTATTTATTATTTATTGTTGA

CCTGCAGGCATGCAAGCTTATCGATACCGTCGACCTCGAGGGGGGGCCCGGTACCCAGCT

TTTGTTCCCTTTAGTGAGGGTTAATTGCGCGCTTGGCGTAATCATGGTCATAGCTGTTTC

CTGTGTGAAATTGTTATCCGCTCACAATTCCACACAACATACGAGCCGGAAGCATAAAGT

GTAAAGCCTGGGGTGCCTAATGAGTGAGCTAACTCACATTAATTGCGTTGCGCTCACTGC

CCGCTTTCCAGTCGGGAAACCTGTCGTGCCAGCTGCATTAATGAATCGGCCAACGCGCGG

GGAGAGGCGGTTTGCGTATTGGGCGCTCTTCCGCTTCCTCGCTCACTGACTCGCTGCGCT

CGGTCGTTCGGCTGCGGCGAGCGGTATCAGCTCACTCAAAGGCGGTAATACGGTTATCCA

CAGAATCAGGGGATAACGCAGGAAAGAACATGTGAGCAAAAGGCCAGCAAAAGGCCAGGA

ACCGTAAAAAGGCCGCGTTGCTGGCGTTTTTCCATAGGCTCCGCCCCCCTGACGAGCATC

ACAAAAATCGACGCTCAAGTCAGAGGTGGCGAAACCCGACAGGACTATAAAGATACCAGG

CGTTTCCCCCTGGAAGCTCCCTCGTGCGCTCTCCTGTTCCGACCCTGCCGCTTACCGGAT

ACCTGTCCGCCTTTCTCCCTTCGGGAAGCGTGGCGCTTTCTCATAGCTCACGCTGTAGGT

ATCTCAGTTCGGTGTAGGTCGTTCGCTCCAAGCTGGGCTGTGTGCACGAACCCCCCGTTC

AGCCCGACCGCTGCGCCTTATCCGGTAACTATCGTCTTGAGTCCAACCCGGTAAGACACG

ACTTATCGCCACTGGCAGCAGCCACTGGTAACAGGATTAGCAGAGCGAGGTATGTAGGCG

GTGCTACAGAGTTCTTGAAGTGGTGGCCTAACTACGGCTACACTAGAAGGACAGTATTTG

GTATCTGCGCTCTGCTGAAGCCAGTTACCTTCGGAAAAAGAGTTGGTAGCTCTTGATCCG

GCAAACAAACCACCGCTGGTAGCGGTGGTTTTTTTGTTTGCAAGCAGCAGATTACGCGCA

GAAAAAAAGGATCTCAAGAAGATCCTTTGATCTTTTCTACGGGGTCTGACGCTCAGTGGA

ACGAAAACTCACGTTAAGGGATTTTGGTCATGAGATTATCAAAAAGGATCTTCACCTAGA

TCCTTTTAAATTAAAAATGAAGTTTTAAATCAATCTAAAGTATATATGAGTAAACTTGGT

CTGACAGTTACCAATGCTTAATCAGTGAGGCACCTATCTCAGCGATCTGTCTATTTCGTT

CATCCATAGTTGCCTGACTCCCCGTCGTGTAGATAACTACGATACGGGAGGGCTTACCAT

CTGGCCCCAGTGCTGCAATGATACCGCGAGACCCACGCTCACCGGCTCCAGATTTATCAG

CAATAAACCAGCCAGCCGGAAGGGCCGAGCGCAGAAGTGGTCCTGCAACTTTATCCGCCT

CCATCCAGTCTATTAATTGTTGCCGGGAAGCTAGAGTAAGTAGTTCGCCAGTTAATAGTT

TGCGCAACGTTGTTGCCATTGCTACAGGCATCGTGGTGTCACGCTCGTCGTTTGGTATGG

CTTCATTCAGCTCCGGTTCCCAACGATCAAGGCGAGTTACATGATCCCCCATGTTGTGCA

AAAAAGCGGTTAGCTCCTTCGGTCCTCCGATCGTTGTCAGAAGTAAGTTGGCCGCAGTGT

TATCACTCATGGTTATGGCAGCACTGCATAATTCTCTTACTGTCATGCCATCCGTAAGAT

GCTTTTCTGTGACTGGTGAGTACTCAACCAAGTCATTCTGAGAATAGTGTATGCGGCGAC

CGAGTTGCTCTTGCCCGGCGTCAATACGGGATAATACCGCGCCACATAGCAGAACTTTAA

AAGTGCTCATCATTGGAAAACGTTCTTCGGGGCGAAAACTCTCAAGGATCTTACCGCTGT

TGAGATCCAGTTCGATGTAACCCACTCGTGCACCCAACTGATCTTCAGCATCTTTTACTT

TCACCAGCGTTTCTGGGTGAGCAAAAACAGGAAGGCAAAATGCCGCAAAAAAGGGAATAA

GGGCGACACGGAAATGTTGAATACTCATACTCTTCCTTTTTCAATATTATTGAAGCATTT

ATCAGGGTTATTGTCTCATGAGCGGATACATATTTGAATGTATTTAGAAAAATAAACAAA

TAGGGGTTCCGCGCACATTTCCCCGAAAAGTGCCAC

Restriction sites to linearize plasmid NdeI

Selectable marker used to select the mutant parasite tgdhfr

Promoter of the selectable marker pbdhfr

Selection (positive) procedure pyrimethamine

Selection (negative) procedure No

Additional remarks genetic modification To generate the transgenic RON4-mCherry, parasites expressing C-terminally tagged mCherry RON4 (PBANKA_932000), construct pL1577 was used (Fougere et al., 2016). The smac (PBANKA_0100600) was exchanged for the ron4 targeting region amplified with primers 6686 and 6687 (5’- gggactagtGCATCCATATTAATGCAACAATGG and 5’-
cccggatccTAAATCATCAAAAATGGCTTTCTC) using restriction sites, SpeI/BamHI. Tranfection (exp. 1964; RMgm-628; www.pberghei.eu) of the linearized pL1837 plasmid (using NdeI), selection and cloning of transformed parasites was performed using standard genetic modification
technologies

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	gggactagtGCATCCATATTAATGCAACAATGG
Additional information primer 1	L6686 (SpeI); PBANKA_093200 tag F
Sequence Primer 2	cccggatccTAAATCATCAAAAATGGCTTTCTC
Additional information primer 2	L6687 (BamHI); PBANKA_093200 tag R
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

top of page