RMgmDB - Rodent Malaria genetically modified Parasites

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Summary

RMgm-1234

Malaria parasite	P. berghei
Genotype
Tagged	Gene model (rodent): PBANKA_1300200; Gene model (P.falciparum): Not available; Gene product: BIR protein PIR protein (PIR protein; PIR5) Name tag: mCherry
Phenotype	Asexual bloodstage;

Last modified: 18 November 2016, 13:03

*RMgm-1234

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) : 27851824
MR4 number
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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).
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The mutant parasite was generated by
Name PI/Researcher	A. Fougère, C.J. Janse, B.M.D. Franke-Fayard
Name Group/Department	Leiden Malaria Research Group
Name Institute	Leiden University Medical Center
City	Leiden
Country	The Netherlands
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Name of the mutant parasite
RMgm number	RMgm-1234
Principal name	2448cl1
Alternative name
Standardized name
Is the mutant parasite cloned after genetic modification	Yes
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Phenotype
Asexual blood stage	The tagged-protein is exported into the cytoplasm of the host erythrocyte and shows a diffuse or patchy localization in the cytoplasm
Gametocyte/Gamete	Not tested
Fertilization and ookinete	Not tested
Oocyst	Not tested
Sporozoite	Not tested
Liver stage	Not tested
Additional remarks phenotype	Mutant/mutation Protein (function) Phenotype Additional information The mutant parasite expresses an mCherry-tagged protein.The protein has been selected for tagging in a screen for putative exported proteins of P. berghei. The genotype of the parasites has not been analysed in detail. Southern analysis of PFG-separated chromosomes (to show integration into the chromosome on which the target gene is located) has been performed (see below). This analysis provided evidence for tagging of the correct gene. The construct used aims at integration of the tagging construct by single-cross-over integration resulting in the presence of a tagged copy of the endogenous gene. Phenotype analyses of the blood stages by fluorescence microscopy show that the tagged-protein is exported into the cytoplasm of the host erythrocyte and shows a diffuse or patchy localization in the cytoplasm. Other mutants

Tagged: Mutant parasite with a tagged gene

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Details of the target gene

Gene Model of Rodent Parasite PBANKA_1300200

Gene Model P. falciparum ortholog Not available

Gene product BIR protein PIR protein

Gene product: Alternative name PIR protein; PIR5

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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 double cross-over

PlasmoGEM (Sanger) construct/vector used No

Modified PlasmoGEM construct/vector used No

Plasmid/construct map

Plasmid/construct sequence

GATCCGGTGGAGGTGGAAGATCCATGGTGAGCAAGGGCGAGGAGGATAACATGGCCATCA

TCAAGGAGTTCATGCGCTTCAAGGTGCACATGGAGGGCTCCGTGAACGGCCACGAGTTCG

AGATCGAGGGCGAGGGCGAGGGCCGCCCCTACGAGGGCACCCAGACCGCCAAGCTGAAGG

TGACCAAGGGTGGCCCCCTGCCCTTCGCCTGGGACATCCTGTCCCCTCAGTTCATGTACG

GCTCCAAGGCCTACGTGAAGCACCCCGCCGACATCCCCGACTACTTGAAGCTGTCCTTCC

CCGAGGGCTTCAAGTGGGAGCGCGTGATGAACTTCGAGGACGGCGGCGTGGTGACCGTGA

CCCAGGACTCCTCCCTGCAGGACGGCGAGTTCATCTACAAGGTGAAGCTGCGCGGCACCA

ACTTCCCCTCCGACGGCCCCGTAATGCAGAAGAAGACCATGGGCTGGGAGGCCTCCTCCG

AGCGGATGTACCCCGAGGACGGCGCCCTGAAGGGCGAGATCAAGCAGAGGCTGAAGCTGA

AGGACGGCGGCCACTACGACGCTGAGGTCAAGACCACCTACAAGGCCAAGAAGCCCGTGC

AGCTGCCCGGCGCCTACAACGTCAACATCAAGTTGGACATCACCTCCCACAACGAGGACT

ACACCATCGTGGAACAGTACGAACGCGCCGAGGGCCGCCACTCCACCGGCGGCATGGACG

AGCTGTACAAGTAATCTAGAATTATTAATATATATGAATATATATACATCGTTGTATGCG

TAATATACGTGTAACTGTTTTTTTTAATTTTTATATTAACTTTTCCTTATACATTTTAAT

GCCATTAACTAATAATATTTTACTTGAAAAAATTCTAAATTTTTATAACTCTAATATAAA

ATCTGTGAATAATAAAAAATTAGCATAAAGTACGAATTAAACATCGAATATTTGTTTTAA

AGGGGCACACATTATGTGCATATAATTTTATTCTTTGAATTTAATTTTTTGCTTATAATT

AGATTAAATTAATATAATTTTTTTATTGAAAATAAAAACTTATTTTAAAAAATTAACAAA

TGGAAGTTAATGAAAAATACGAAAAAAGTAAAGGGTTAATTCTTATATGGTCGGTACCGA

GCTCGAATTCGATATCGAAATTGAAGGAAAAAACATCATTTGTGTTTATATGTTTTTTTT

TAATTTTTCAACTTTTTTTATTTAACTATGCTAATCATATTATAAACATTTTAAAAATCA

AGTCCAACTATTTATGAATCATTGAAGAGACAACATAATTATAACAGTAAAAATAATAAT

AATAATAATAATTTTGTCTTATTGTTTCAAACATATTATGTTCCATAAATATCTAAATTT

TTAATATGGTCATAAATTCATAGGATTTAATTTATATAAAATATGGCTATTCATACTAGC

CATTTTATGTGTGTAAAGTTTTTTTTTTCTGAACAAAAAATAAAATGTTCATAATTTTAG

CTATTTACATGCATGTGCATGCACAAAAAAAAATATGCACACAACATACACATTTTTACA

GTTATAAATACAATCAATTGGTATAAATATATAAGTAAGAAAAACGGGATCAAAGCTAGC

TTCTGTATTTCCGCTAGACAGCCATCTCCATCTGGATTCGTCCGTGCGGGACGTAGCCCA

CGACCTCAAAATCCTCGGCGGTGAAATCGTCGATTTCCTTGATGCGTTCCTTGTTGAGGA

TGTTCACAATGGGGAACGGTCTCGGTTCTCTCCGCAGCTGCTCTTTTAAAGCCTCGACAT

GGTTCGTGTAGACATGCGTGTTCCCCATGAAGTGAATGAACTCCTTAGGTTTTAGGTTGC

AGACGTGTGCAACCATGAGCGTCAAAAGCGAATAGGAAGCGATGTTGAAGGGGACGCCGA

GGCCGACATCGCACGACCGCTGATACATGATGCACGACAGCTCCTTCTGGTCGTTCACGT

AGAACTGGCACAACAAGTGACAAGGCGGCAGCGCCATTTCGTCCAGCGCTGCAGGATTCC

AGGCAGTCATGAGCATGCGACGATCTGTTGGATTCGTTCTCAGCATCTGGATCACATTCT

TCAGCTGGTCGACGCCCTGCCCTGTGTAGTCTGTGTGCATGTCTTTGTATGCCGCGCCGA

AGTGTCTCCACTGGAAGCCGTAGCCCGGGCCGATGTCTCCGACCTCTCGGTGGGGGAGAT

TGCGCGAATCGAGGAACTCGCGTGTCACATTCTTGTCCCAGATCTTCACGCCCTTCTCAG

AAAGATGGTTTGCGTTCGTGTCGCCGCGAATGAACCACAGCAACTCTTCGAGGACCCCTT

TCCAGAACACACGCTTTGTGGTGAGAAGTGGAAAGGCCTGATCCAGCGAGTAGCGCATAG

TGCAGCCGAATTTGGAGATGACACCAACGCCCGTTCGGTCATCCATTGTCCTTCCATTGT

TAATAATGTCGGCAATGAGATCAAGGTACTGGAACTCTTCATGGCCTCTAAAGTGAACAT

GCGGAACGGCCCGAATCAGTTCCTTTTGCTCGCGTTTTTTCCGGTCTTCTTCGTCCATCC

ACGCCAACACCGGGGCAATGGCTGCGGCCGAAGAAGGAGCCTGCAACCCGTGCACGGGAG

TTGTCTCCCTCGTGGACGTCAAGGAGCTCATTGCGTTGCTCGGTTCCGCAGTGGCTGCGT

CGTCAGTCTTCCTTCTCTTCTCGAGAACCACAAAGTCGTAGGGAACCCCGTTGTCTGAGA

AGGTCTTGGAAATGAAGATGGGTCGATACGTCGCTTCATTGTCCTTCTCTCTTCCGAGCT

CCGGACAAAAGGGAACGAACACAGACTCGGCAGGAGCTGCAGCCTGCGCAGCAGTTGATT

TGTTTGAAAGAATGTCATCTCCGGGGAACGCAGGGAAGAAAACGTCGCACGGAAACTCGC

GGGCTACACGCGTGATGTACAGGTGAGAGGCAACGCCCAGAGACAGCGCTGCCTCGTACA

GTCCCGCTCCTCCCACGACAAAAATCTGGTCGACAGAATCCTTGTACTCTTCCTCCAGAA

GGCTGAGAGCTGCTGGGAGTGAAGCACAGACTCGGACGCGCTGCTGGCCTTCAGCTTGAG

GCTTCTCCGCCGCAATGTCTTCTTCTTTGAGGGAAGAGGAAACGACGATGTTCAATCTGT

CCACGAGGGGTCTAAACTTTCGAGGCATGCTTTCCCAGGTTTTCCGTCCCATGACAACGG

CGTTGAATCTCTTGCCGACTGATGGAGAGGGAAGTCCAGAGTCGCCCGTCTTTGCAAATT

TCCTGGGAAGCCACCCGTTCAGGCGACTGGCTTCTTCGGGCGTCGTTTTTGTCACACGAG

AAAAGTGTTTGAAATCTGTGGTCAAGTGGGGCCACGGGAGGCCGTTGTTGATGCCGATGC

CCCTCTTGGGGGTCATCGCGACGACCAGACACACCGGTTTATGCATTGATCCTTTGTAAC

ATTTAGGTGTGTATTTATATATATAAGCAATATACAAATGAAAATACTATTATAAAACGG

AAACTCAAAAAATGTTGAAACAAATAGTCAATTTTTAAAATGAATGTGTAAATTGTTTAA

GTTTTATGACAAAAAAAGGAAAAGTGTATATAAAATAAAAATTGGAAAAAATTTATATAT

TACATGCCATATAATACCATTTATTTACTATTTATATCTTAATATAAAAAATTGTAGCAA

AAATATATAATCGTATTTTATATATTTAATATTATATAACCCCTTAAAAAAAAAAATTTA

ATTGAAAAAATTTTAATTTATGCACTTTTTTATTGTATTATATATATTCCGTTATTTTAA

TTACACATAAACTTAATAGAAAAATTAATTCGCTGCTAAATAAAAATTATAGGGCTATAT

ATGATGGGATCATATTCTATCTTTCTCTTTTTCAAATYCTTTATATTTTAGAATACAAAT

CGCTTTAAACAATAAAAAAGAGGTATATCCTACCCGAATTGACACACACAATAAATAATA

AATATATAAATAATAAAAATATAAATATATAAATTATAAATATATAAATTATATATATAA

ATATATTAACATGTGTGTAATTTGTTAAAATTGATATTAAACAGCTTTGCATGGGGAAAG

AGTGGCTTTTATAAGCGAATCATATAATACAGTGTTAATTTTAATATATAAAATATAATT

TCCTTAAATTGTTATTTAACATTTTTAATTGCCTACCCARGGTAATATAATATCCAAATG

ACAATCAAAGATAAATGAAATACCGCTCCATTTTTCCTATATATCCATTTTCATTTTGTG

TAAATGTTCATAAAATTTTATCATATACTCATAAAAAACAGGGGAAAAATGAAAAAATGA

ACGTCCACCCCGTGTGAATATGCTCATTTTGTATACTATACATTTTTTTTTATATAATTT

ATATTTTCCAAATAAAAAAAATGTTTGTACATGTATATATCAATATTGTGTTATGGCATG

ATGAAAAACAATTATTTTATTCGTTTTTAAATTTGTAGAATTTCCCTAAATTATTAATGA

TTTCATATTATAATATATGTATCTATATATTTTTAATGTGTAATTCAAAATGGCGAAAAA

GAAACAAACCATACTTTATAAGCAACTCTCTTTGAGCACTTACCATAAAAAATTATACTA

CTAAATTCAAATTAAATAAAAAAATATATATTATAAAATTGGCTTTAATTATTTATAAGC

ACTTGCTTTGTTTTACTCAAAAAGTATTGGCATAAGTATAAACATAAAACCAATTTTTCA

CAAAATATATTATATGTCCATACAACTATATCCGAACYTTCAAAACACATCCAAATTTCC

ATTTACCACATGCAAGCAAAAGGATATTAAATAAAAAAAAATTATTTATATTTTCATGAT

CACTTAAATAATATTTAAAAAAAAAAGAGACCATATATAAAATGTATAATTTAAATATAG

CTAACTTATTAAAAAAAATGAATATTGAGCATATATCCATAATTTAAAACACTTCTATTT

TATGTTATTTGATATATAAGTCGATAAATTATTGCATGTATGCATATAGACAGATTAATA

TACACCAAAATAATAAAAATTATATATAGTTCATATATTATTCTGAATAATAAATTAGCT

TTCCTCTTAATTCACTTGGTATGTATATATATATATATATATATATATATATATATATGG

ATAAATAAATATAAGGTCTATAACAGGAAAAAATGTGTATTAACTTTTTTTTATAATTAT

TTTATATTTTTTCTCCATTTATTTAAATATGTTAAAATTATTTCCCATTTTATAAACAAA

AATTAATCCATAAAATAGCAAAATAACAAAATGACAAAATAACAAAAACAAAAAAACGCA

TTATATGAGTTCATTTTACACAATCCAAAGAAAAAGAAAATATGCTTAAAATATTCATAA

CACACTTTTAAGCTATAAAAATATATATATTTATTTGTGTCTATATTACCAACTCAATTT

AATAGATGTGTTATGTGATTAATTCATACACAAACATACAAAAATAAACACCATTTTTGA

AAAGATTAATTTGAAATCATATATGTTATTATTATATATTTTTATATCAATGAAATTTAA

AAAAATAAAAGGGAAATCAATGTATTAAAAATAATTATATGTTATTTTATTTCCACAATA

TTTATTTATTATTTATTGTTGACCTGCAGGCATGCAAGCTTCTGGATAATTAATATATGA

CCATGAGGATAGCGATCAATATACTTTAAGATATTTTTATATTTGGAAATAACAAAATTT

TGATCATAATTTTTGCATAATTTTTATATAATTTTTATGTTATGAGTCAGGGTTGAGGTT

ATGTTTGTAGAACCCATATTCGGGTTAGGATTAAATGTTACATTGTATTTAATTTTGTAT

AATTTTATAATATTTATTGATATGTATGTGTTGATCAAATATATTTACAATTTCCGTATG

CTTAATCTCAAGCTGAAGTCTAAATATGCAACCAAAAAAGGGGCATTTCCATTAATATGA

AAAGGGCCATAAACATTTTTTCATAAGTTATAATATATATAATTAATGTTTATGTCTATT

TAATATGATTAAAAAAATGTCTATATTGCATATATTAATTCATGATATATCATACTTATT

TATTATATAAAATTTGTTAATCTGTGGTTATATCGAATTATATATAACACAACATGTTTC

TTTATGTGATGAAACATTTTGTTTAGTAATACCGCGGCCGCGGGCCCGGTACCCAGCTTT

TGTTCCCTTTAGTGAGGGTTAATTGCGCGCTTGGCGTAATCATGGTCATAGCTGTTTCCT

GTGTGAAATTGTTATCCGCTCACAATTCCACACAACATACGAGCCGGAAGCATAAAGTGT

AAAGCCTGGGGTGCCTAATGAGTGAGCTAACTCACATTAATTGCGTTGCGCTCACTGCCC

GCTTTCCAGTCGGGAAACCTGTCGTGCCAGCTGCATTAATGAATCGGCCAACGCGCGGGG

AGAGGCGGTTTGCGTATTGGGCGCTCTTCCGCTTCCTCGCTCACTGACTCGCTGCGCTCG

GTCGTTCGGCTGCGGCGAGCGGTATCAGCTCACTCAAAGGCGGTAATACGGTTATCCACA

GAATCAGGGGATAACGCAGGAAAGAACATGTGAGCAAAAGGCCAGCAAAAGGCCAGGAAC

CGTAAAAAGGCCGCGTTGCTGGCGTTTTTCCATAGGCTCCGCCCCCCTGACGAGCATCAC

AAAAATCGACGCTCAAGTCAGAGGTGGCGAAACCCGACAGGACTATAAAGATACCAGGCG

TTTCCCCCTGGAAGCTCCCTCGTGCGCTCTCCTGTTCCGACCCTGCCGCTTACCGGATAC

CTGTCCGCCTTTCTCCCTTCGGGAAGCGTGGCGCTTTCTCATAGCTCACGCTGTAGGTAT

CTCAGTTCGGTGTAGGTCGTTCGCTCCAAGCTGGGCTGTGTGCACGAACCCCCCGTTCAG

CCCGACCGCTGCGCCTTATCCGGTAACTATCGTCTTGAGTCCAACCCGGTAAGACACGAC

TTATCGCCACTGGCAGCAGCCACTGGTAACAGGATTAGCAGAGCGAGGTATGTAGGCGGT

GCTACAGAGTTCTTGAAGTGGTGGCCTAACTACGGCTACACTAGAAGGACAGTATTTGGT

ATCTGCGCTCTGCTGAAGCCAGTTACCTTCGGAAAAAGAGTTGGTAGCTCTTGATCCGGC

AAACAAACCACCGCTGGTAGCGGTGGTTTTTTTGTTTGCAAGCAGCAGATTACGCGCAGA

AAAAAAGGATCTCAAGAAGATCCTTTGATCTTTTCTACGGGGTCTGACGCTCAGTGGAAC

GAAAACTCACGTTAAGGGATTTTGGTCATGAGATTATCAAAAAGGATCTTCACCTAGATC

CTTTTAAATTAAAAATGAAGTTTTAAATCAATCTAAAGTATATATGAGTAAACTTGGTCT

GACAGTTACCAATGCTTAATCAGTGAGGCACCTATCTCAGCGATCTGTCTATTTCGTTCA

TCCATAGTTGCCTGACTCCCCGTCGTGTAGATAACTACGATACGGGAGGGCTTACCATCT

GGCCCCAGTGCTGCAATGATACCGCGAGACCCACGCTCACCGGCTCCAGATTTATCAGCA

ATAAACCAGCCAGCCGGAAGGGCCGAGCGCAGAAGTGGTCCTGCAACTTTATCCGCCTCC

ATCCAGTCTATTAATTGTTGCCGGGAAGCTAGAGTAAGTAGTTCGCCAGTTAATAGTTTG

CGCAACGTTGTTGCCATTGCTACAGGCATCGTGGTGTCACGCTCGTCGTTTGGTATGGCT

TCATTCAGCTCCGGTTCCCAACGATCAAGGCGAGTTACATGATCCCCCATGTTGTGCAAA

AAAGCGGTTAGCTCCTTCGGTCCTCCGATCGTTGTCAGAAGTAAGTTGGCCGCAGTGTTA

TCACTCATGGTTATGGCAGCACTGCATAATTCTCTTACTGTCATGCCATCCGTAAGATGC

TTTTCTGTGACTGGTGAGTACTCAACCAAGTCATTCTGAGAATAGTGTATGCGGCGACCG

AGTTGCTCTTGCCCGGCGTCAATACGGGATAATACCGCGCCACATAGCAGAACTTTAAAA

GTGCTCATCATTGGAAAACGTTCTTCGGGGCGAAAACTCTCAAGGATCTTACCGCTGTTG

AGATCCAGTTCGATGTAACCCACTCGTGCACCCAACTGATCTTCAGCATCTTTTACTTTC

ACCAGCGTTTCTGGGTGAGCAAAAACAGGAAGGCAAAATGCCGCAAAAAAGGGAATAAGG

GCGACACGGAAATGTTGAATACTCATACTCTTCCTTTTTCAATATTATTGAAGCATTTAT

CAGGGTTATTGTCTCATGAGCGGATACATATTTGAATGTATTTAGAAAAATAAACAAATA

GGGGTTCCGCGCACATTTCCCCGAAAAGTGCCACCTAAATTGTAAGCGTTAATATTTTGT

TAAAATTCGCGTTAAATTTTTGTTAAATCAGCTCATTTTTTAACCAATAGGCCGAAATCG

GCAAAATCCCTTATAAATCAAAAGAATAGACCGAGATAGGGTTGAGTGTTGTTCCAGTTT

GGAACAAGAGTCCACTATTAAAGAACGTGGACTCCAACGTCAAAGGGCGAAAAACCGTCT

ATCAGGGCGATGGCCCACTACGTGAACCATCACCCTAATCAAGTTTTTTGGGGTCGAGGT

GCCGTAAAGCACTAAATCGGAACCCTAAAGGGAGCCCCCGATTTAGAGCTTGACGGGGAA

AGCCGGCGAACGTGGCGAGAAAGGAAGGGAAGAAAGCGAAAGGAGCGGGCGCTAGGGCGC

TGGCAAGTGTAGCGGTCACGCTGCGCGTAACCACCACACCCGCCGCGCTTAATGCGCCGC

TACAGGGCGCGTCCCATTCGCCATTCAGGCTGCGCAACTGTTGGGAAGGGCGATCGGTGC

GGGCCTCTTCGCTATTACGCCAGCTGGCGAAAGGGGGATGTGCTGCAAGGCGATTAAGTT

GGGTAACGCCAGGGTTTTCCCAGTCACGACGTTGTAAAACGACGGCCAGTGAGCGCGCGT

AATACGACTCACTATAGGGCGAATTGGAGCTCCACCGCGGTGGCGGCCGCTCTAGAACTA

GTGAACACTATGGCTATGTTGTATGATGCATTTAAAATTTTATGTAATATGTATACTGAG

TTTGACGAAAAAAAGCTAAATTGCGATGAATGTTTGAAATATGGTGATGAATTTGTTGAA

AAATATGATCATCTTAATAAAATTTCTGAAATTACTGAAGGCAGTCCATACTATCAAGTA

TTATCTACTTTATCAAATGATTATGATAATTTTAAAAAGAAATATAATGAAGCTCAATGC

TGCAATTTTCCACCTCTTCCAACGATAGAAAAAGCACAAGCTTCTGCACAAAGTTCTAAA

TTTACATCATCAAGTTCGTCGATAGAAAGAAAATTATTTATAGTTTTATCGATATTTGGT

GCAATAGCATTTTTTTTTGGAATTTCTTATAAGGTAAATAATAAGGAATTTAAAAATAAT

TTTCATTATATATATGTAAACATTAACAAAAATATCATATGCTTCTTAACATTTTATATT

AGTATTCGTTATTTGAATTTCGGAAACGATCTCAAAAACAACATTTAAGAGAAAAGCTAA

AAAAAG

Restriction sites to linearize plasmid NotI, ScaI

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

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

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