RMgmDB - Rodent Malaria genetically modified Parasites

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Summary

RMgm-714

Malaria parasite	P. berghei
Genotype
Tagged	Gene model (rodent): PBANKA_1238800; Gene model (P.falciparum): PF3D7_0524000; Gene product: karyopherin beta (KASbeta) Name tag: mCherry
Phenotype	Asexual bloodstage;

Last modified: 2 December 2012, 15:42

*RMgm-714

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) : 23197789
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	E.M. Pasini, C.J. Janse, B.M.D. Franke-Fayard
Name Group/Department	Department of Parasitology
Name Institute	Biomedical Primate Research Centre
City	Rijswijk
Country	The Netherlands
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Name of the mutant parasite
RMgm number	RMgm-714
Principal name	1956
Alternative name
Standardized name
Is the mutant parasite cloned after genetic modification	No
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Phenotype
Asexual blood stage	The tagged-protein shows a predominant location in the cytoplasm of the parasite
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 analyzed 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 has a predominant location in the cytoplasm of the parasite. Other mutants

Tagged: Mutant parasite with a tagged gene

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Details of the target gene

Gene Model of Rodent Parasite PBANKA_1238800

Gene Model P. falciparum ortholog PF3D7_0524000

Gene product karyopherin beta

Gene product: Alternative name KASbeta

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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 Clontech

Type of plasmid/construct 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

CCGCGGTGGCGGCCGCTCTAGAACTAGTCAGCAAGAATAATATCAGATCAAACAGATAAT

AATAAATCTATGCTATTAGCTATACTTACAACATCTGCTGAAAAAGTATTAAAAAGTTTA

TCAGATACAAAATTAGATGATAATTATGAATATGTATTAGATATAATGATTATCGAATCT

AATGGTTTATATATGTGCTTACAAAAAGCCGGTGCTAATATATTACCTAACAATACATTA

AAAATGTTTTTTAATGAAATATTTAAATTATTACAATATTCAACAGATAGAAGAGTCATT

TATAATCAAAAAAAAAATAATGAAGATGTCGATGATGATGAATTATTAATTATAGACAGA

GAAGAAGAGCTAGAACAAACATATCGAACAAACTTATTAGACATATTGGGAGTATTAATA

AAACATCACACAAATCAATTCTTAAGTACATGTTGTGATATATGTATTACATTTATAAAT

AATTATTTAAACTCACCACATGCAGAAGATATTGCATTGGCTTTATATGTTTGTGATGAT

TTGTTAGAATTTTTGCAAGATAATAGTGTATGCTTGTGGGAATATTTTATGAACCCATTA

TTATTAAATATTAATCATACAGATCATAAAGTTAAACAAGCAGCTTGTTATGGTGTTATA

CAAGCAAATAAAATAGAAGCATTTGGAAAATATGCAAATTTGGCCATAGAATATTTATTA

AAACTATTACATCAAACTCCCCCAAGTAAAAAACCCAAGGAATACATATCTGCTATTGAT

AATGCTGTAGCTGCATTAGGAGATGTTGTCTTAATGCACACATCCAAATTCAATAATTCA

GAAGAATTAATTAAGTTGTGGTTAAATAATTTACCAATTAAAGAAGATGAATCAGAAGGA

AGACGAGTGCACAAAAACTTAATCGATTTAGTTTCTCAAAATCACCCACTTTTATTCGGA

AAAGATAATTCTAATATTGGAAAAATTATAGAAATATTCCTATCAATATATGAAACAGAA

TTTTCTGATTCAGATTGTAACAAAAAAATAGTATCCTTGATAAGTTCTTTAGATCAATCT

TATTTAAGCAATTTGGCTTCCACATCTCTCACCAACAAACAATCCAAAAAATTGAATCAC

ATTATGAACGCCTCTAGAAAGGGATCCGGTGGAGGTGGAAGATCCATGGTGAGCAAGGGC

GAGGAGGATAACATGGCCATCATCAAGGAGTTCATGCGCTTCAAGGTGCACATGGAGGGC

TCCGTGAACGGCCACGAGTTCGAGATCGAGGGCGAGGGCGAGGGCCGCCCCTACGAGGGC

ACCCAGACCGCCAAGCTGAAGGTGACCAAGGGTGGCCCCCTGCCCTTCGCCTGGGACATC

CTGTCCCCTCAGTTCATGTACGGCTCCAAGGCCTACGTGAAGCACCCCGCCGACATCCCC

GACTACTTGAAGCTGTCCTTCCCCGAGGGCTTCAAGTGGGAGCGCGTGATGAACTTCGAG

GACGGCGGCGTGGTGACCGTGACCCAGGACTCCTCCCTGCAGGACGGCGAGTTCATCTAC

AAGGTGAAGCTGCGCGGCACCAACTTCCCCTCCGACGGCCCCGTAATGCAGAAGAAGACC

ATGGGCTGGGAGGCCTCCTCCGAGCGGATGTACCCCGAGGACGGCGCCCTGAAGGGCGAG

ATCAAGCAGAGGCTGAAGCTGAAGGACGGCGGCCACTACGACGCTGAGGTCAAGACCACC

TACAAGGCCAAGAAGCCCGTGCAGCTGCCCGGCGCCTACAACGTCAACATCAAGTTGGAC

ATCACCTCCCACAACGAGGACTACACCATCGTGGAACAGTACGAACGCGCCGAGGGCCGC

CACTCCACCGGCGGCATGGACGAGCTGTACAAGTAATCTAGAATTATTAATATATATGAA

TATATATACATCGTTGTATGCGTAATATACGTGTAACTGTTTTTTTTAATTTTTATATTA

ACTTTTCCTTATACATTTTAATGCCATTAACTAATAATATTTTACTTGAAAAAATTCTAA

ATTTTTATAACTCTAATATAAAATCTGTGAATAATAAAAAATTAGCATAAAGTACGAATT

AAACATCGAATATTTGTTTTAAAGGGGCACACATTATGTGCATATAATTTTATTCTTTGA

ATTTAATTTTTTGCTTATAATTAGATTAAATTAATATAATTTTTTTATTGAAAATAAAAA

CTTATTTTAAAAAATTAACAAATGGAAGTTAATGAAAAATACGAAAAAAGTAAAGGGTTA

ATTCTTATATGGTCGGTACCGAGCTCGAATTCGATATCGAAATTGAAGGAAAAAACATCA

TTTGTGTTTATATGTTTTTTTTTAATTTTTCAACTTTTTTTATTTAACTATGCTAATCAT

ATTATAAACATTTTAAAAATCAAGTCCAACTATTTATGAATCATTGAAGAGACAACATAA

TTATAACAGTAAAAATAATAATAATAATAATAATTTTGTCTTATTGTTTCAAACATATTA

TGTTCCATAAATATCTAAATTTTTAATATGGTCATAAATTCATAGGATTTAATTTATATA

AAATATGGCTATTCATACTAGCCATTTTATGTGTGTAAAGTTTTTTTTTTCTGAACAAAA

AATAAAATGTTCATAATTTTAGCTATTTACATGCATGTGCATGCACAAAAAAAAATATGC

ACACAACATACACATTTTTACAGTTATAAATACAATCAATTGGTATAAATATATAAGTAA

GAAAAACGGGATCAAAGCTAGCTTCTGTATTTCCGCTAGACAGCCATCTCCATCTGGATT

CGTCCGTGCGGGACGTAGCCCACGACCTCAAAATCCTCGGCGGTGAAATCGTCGATTTCC

TTGATGCGTTCCTTGTTGAGGATGTTCACAATGGGGAACGGTCTCGGTTCTCTCCGCAGC

TGCTCTTTTAAAGCCTCGACATGGTTCGTGTAGACATGCGTGTTCCCCATGAAGTGAATG

AACTCCTTAGGTTTTAGGTTGCAGACGTGTGCAACCATGAGCGTCAAAAGCGAATAGGAA

GCGATGTTGAAGGGGACGCCGAGGCCGACATCGCACGACCGCTGATACATGATGCACGAC

AGCTCCTTCTGGTCGTTCACGTAGAACTGGCACAACAAGTGACAAGGCGGCAGCGCCATT

TCGTCCAGCGCTGCAGGATTCCAGGCAGTCATGAGCATGCGACGATCTGTTGGATTCGTT

CTCAGCATCTGGATCACATTCTTCAGCTGGTCGACGCCCTGCCCTGTGTAGTCTGTGTGC

ATGTCTTTGTATGCCGCGCCGAAGTGTCTCCACTGGAAGCCGTAGCCCGGGCCGATGTCT

CCGACCTCTCGGTGGGGGAGATTGCGCGAATCGAGGAACTCGCGTGTCACATTCTTGTCC

CAGATCTTCACGCCCTTCTCAGAAAGATGGTTTGCGTTCGTGTCGCCGCGAATGAACCAC

AGCAACTCTTCGAGGACCCCTTTCCAGAACACACGCTTTGTGGTGAGAAGTGGAAAGGCC

TGATCCAGCGAGTAGCGCATAGTGCAGCCGAATTTGGAGATGACACCAACGCCCGTTCGG

TCATCCATTGTCCTTCCATTGTTAATAATGTCGGCAATGAGATCAAGGTACTGGAACTCT

TCATGGCCTCTAAAGTGAACATGCGGAACGGCCCGAATCAGTTCCTTTTGCTCGCGTTTT

TTCCGGTCTTCTTCGTCCATCCACGCCAACACCGGGGCAATGGCTGCGGCCGAAGAAGGA

GCCTGCAACCCGTGCACGGGAGTTGTCTCCCTCGTGGACGTCAAGGAGCTCATTGCGTTG

CTCGGTTCCGCAGTGGCTGCGTCGTCAGTCTTCCTTCTCTTCTCGAGAACCACAAAGTCG

TAGGGAACCCCGTTGTCTGAGAAGGTCTTGGAAATGAAGATGGGTCGATACGTCGCTTCA

TTGTCCTTCTCTCTTCCGAGCTCCGGACAAAAGGGAACGAACACAGACTCGGCAGGAGCT

GCAGCCTGCGCAGCAGTTGATTTGTTTGAAAGAATGTCATCTCCGGGGAACGCAGGGAAG

AAAACGTCGCACGGAAACTCGCGGGCTACACGCGTGATGTACAGGTGAGAGGCAACGCCC

AGAGACAGCGCTGCCTCGTACAGTCCCGCTCCTCCCACGACAAAAATCTGGTCGACAGAA

TCCTTGTACTCTTCCTCCAGAAGGCTGAGAGCTGCTGGGAGTGAAGCACAGACTCGGACG

CGCTGCTGGCCTTCAGCTTGAGGCTTCTCCGCCGCAATGTCTTCTTCTTTGAGGGAAGAG

GAAACGACGATGTTCAATCTGTCCACGAGGGGTCTAAACTTTCGAGGCATGCTTTCCCAG

GTTTTCCGTCCCATGACAACGGCGTTGAATCTCTTGCCGACTGATGGAGAGGGAAGTCCA

GAGTCGCCCGTCTTTGCAAATTTCCTGGGAAGCCACCCGTTCAGGCGACTGGCTTCTTCG

GGCGTCGTTTTTGTCACACGAGAAAAGTGTTTGAAATCTGTGGTCAAGTGGGGCCACGGG

AGGCCGTTGTTGATGCCGATGCCCCTCTTGGGGGTCATCGCGACGACCAGACACACCGGT

TTATGCATTGATCCTTTGTAACATTTAGGTGTGTATTTATATATATAAGCAATATACAAA

TGAAAATACTATTATAAAACGGAAACTCAAAAAATGTTGAAACAAATAGTCAATTTTTAA

AATGAATGTGTAAATTGTTTAAGTTTTATGACAAAAAAAGGAAAAGTGTATATAAAATAA

AAATTGGAAAAAATTTATATATTACATGCCATATAATACCATTTATTTACTATTTATATC

TTAATATAAAAAATTGTAGCAAAAATATATAATCGTATTTTATATATTTAATATTATATA

ACCCCTTAAAAAAAAAAATTTAATTGAAAAAATTTTAATTTATGCACTTTTTTATTGTAT

TATATATATTCCGTTATTTTAATTACACATAAACTTAATAGAAAAATTAATTCGCTGCTA

AATAAAAATTATAGGGCTATATATGATGGGATCATATTCTATCTTTCTCTTTTTCAAATY

CTTTATATTTTAGAATACAAATCGCTTTAAACAATAAAAAAGAGGTATATCCTACCCGAA

TTGACACACACAATAAATAATAAATATATAAATAATAAAAATATAAATATATAAATTATA

AATATATAAATTATATATATAAATATATTAACATGTGTGTAATTTGTTAAAATTGATATT

AAACAGCTTTGCATGGGGAAAGAGTGGCTTTTATAAGCGAATCATATAATACAGTGTTAA

TTTTAATATATAAAATATAATTTCCTTAAATTGTTATTTAACATTTTTAATTGCCTACCC

ARGGTAATATAATATCCAAATGACAATCAAAGATAAATGAAATACCGCTCCATTTTTCCT

ATATATCCATTTTCATTTTGTGTAAATGTTCATAAAATTTTATCATATACTCATAAAAAA

CAGGGGAAAAATGAAAAAATGAACGTCCACCCCGTGTGAATATGCTCATTTTGTATACTA

TACATTTTTTTTTATATAATTTATATTTTCCAAATAAAAAAAATGTTTGTACATGTATAT

ATCAATATTGTGTTATGGCATGATGAAAAACAATTATTTTATTCGTTTTTAAATTTGTAG

AATTTCCCTAAATTATTAATGATTTCATATTATAATATATGTATCTATATATTTTTAATG

TGTAATTCAAAATGGCGAAAAAGAAACAAACCATACTTTATAAGCAACTCTCTTTGAGCA

CTTACCATAAAAAATTATACTACTAAATTCAAATTAAATAAAAAAATATATATTATAAAA

TTGGCTTTAATTATTTATAAGCACTTGCTTTGTTTTACTCAAAAAGTATTGGCATAAGTA

TAAACATAAAACCAATTTTTCACAAAATATATTATATGTCCATACAACTATATCCGAACY

TTCAAAACACATCCAAATTTCCATTTACCACATGCAAGCAAAAGGATATTAAATAAAAAA

AAATTATTTATATTTTCATGATCACTTAAATAATATTTAAAAAAAAAAGAGACCATATAT

AAAATGTATAATTTAAATATAGCTAACTTATTAAAAAAAATGAATATTGAGCATATATCC

ATAATTTAAAACACTTCTATTTTATGTTATTTGATATATAAGTCGATAAATTATTGCATG

TATGCATATAGACAGATTAATATACACCAAAATAATAAAAATTATATATAGTTCATATAT

TATTCTGAATAATAAATTAGCTTTCCTCTTAATTCACTTGGTATGTATATATATATATAT

ATATATATATATATATATATGGATAAATAAATATAAGGTCTATAACAGGAAAAAATGTGT

ATTAACTTTTTTTTATAATTATTTTATATTTTTTCTCCATTTATTTAAATATGTTAAAAT

TATTTCCCATTTTATAAACAAAAATTAATCCATAAAATAGCAAAATAACAAAATGACAAA

ATAACAAAAACAAAAAAACGCATTATATGAGTTCATTTTACACAATCCAAAGAAAAAGAA

AATATGCTTAAAATATTCATAACACACTTTTAAGCTATAAAAATATATATATTTATTTGT

GTCTATATTACCAACTCAATTTAATAGATGTGTTATGTGATTAATTCATACACAAACATA

CAAAAATAAACACCATTTTTGAAAAGATTAATTTGAAATCATATATGTTATTATTATATA

TTTTTATATCAATGAAATTTAAAAAAATAAAAGGGAAATCAATGTATTAAAAATAATTAT

ATGTTATTTTATTTCCACAATATTTATTTATTATTTATTGTTGACCTGCAGGCATGCAAG

CTTATCGATACCGTCGACCTCGAGGGGGGGCCCGGTACCCAGCTTTTGTTCCCTTTAGTG

AGGGTTAATTGCGCGCTTGGCGTAATCATGGTCATAGCTGTTTCCTGTGTGAAATTGTTA

TCCGCTCACAATTCCACACAACATACGAGCCGGAAGCATAAAGTGTAAAGCCTGGGGTGC

CTAATGAGTGAGCTAACTCACATTAATTGCGTTGCGCTCACTGCCCGCTTTCCAGTCGGG

AAACCTGTCGTGCCAGCTGCATTAATGAATCGGCCAACGCGCGGGGAGAGGCGGTTTGCG

TATTGGGCGCTCTTCCGCTTCCTCGCTCACTGACTCGCTGCGCTCGGTCGTTCGGCTGCG

GCGAGCGGTATCAGCTCACTCAAAGGCGGTAATACGGTTATCCACAGAATCAGGGGATAA

CGCAGGAAAGAACATGTGAGCAAAAGGCCAGCAAAAGGCCAGGAACCGTAAAAAGGCCGC

GTTGCTGGCGTTTTTCCATAGGCTCCGCCCCCCTGACGAGCATCACAAAAATCGACGCTC

AAGTCAGAGGTGGCGAAACCCGACAGGACTATAAAGATACCAGGCGTTTCCCCCTGGAAG

CTCCCTCGTGCGCTCTCCTGTTCCGACCCTGCCGCTTACCGGATACCTGTCCGCCTTTCT

CCCTTCGGGAAGCGTGGCGCTTTCTCATAGCTCACGCTGTAGGTATCTCAGTTCGGTGTA

GGTCGTTCGCTCCAAGCTGGGCTGTGTGCACGAACCCCCCGTTCAGCCCGACCGCTGCGC

CTTATCCGGTAACTATCGTCTTGAGTCCAACCCGGTAAGACACGACTTATCGCCACTGGC

AGCAGCCACTGGTAACAGGATTAGCAGAGCGAGGTATGTAGGCGGTGCTACAGAGTTCTT

GAAGTGGTGGCCTAACTACGGCTACACTAGAAGGACAGTATTTGGTATCTGCGCTCTGCT

GAAGCCAGTTACCTTCGGAAAAAGAGTTGGTAGCTCTTGATCCGGCAAACAAACCACCGC

TGGTAGCGGTGGTTTTTTTGTTTGCAAGCAGCAGATTACGCGCAGAAAAAAAGGATCTCA

AGAAGATCCTTTGATCTTTTCTACGGGGTCTGACGCTCAGTGGAACGAAAACTCACGTTA

AGGGATTTTGGTCATGAGATTATCAAAAAGGATCTTCACCTAGATCCTTTTAAATTAAAA

ATGAAGTTTTAAATCAATCTAAAGTATATATGAGTAAACTTGGTCTGACAGTTACCAATG

CTTAATCAGTGAGGCACCTATCTCAGCGATCTGTCTATTTCGTTCATCCATAGTTGCCTG

ACTCCCCGTCGTGTAGATAACTACGATACGGGAGGGCTTACCATCTGGCCCCAGTGCTGC

AATGATACCGCGAGACCCACGCTCACCGGCTCCAGATTTATCAGCAATAAACCAGCCAGC

CGGAAGGGCCGAGCGCAGAAGTGGTCCTGCAACTTTATCCGCCTCCATCCAGTCTATTAA

TTGTTGCCGGGAAGCTAGAGTAAGTAGTTCGCCAGTTAATAGTTTGCGCAACGTTGTTGC

CATTGCTACAGGCATCGTGGTGTCACGCTCGTCGTTTGGTATGGCTTCATTCAGCTCCGG

TTCCCAACGATCAAGGCGAGTTACATGATCCCCCATGTTGTGCAAAAAAGCGGTTAGCTC

CTTCGGTCCTCCGATCGTTGTCAGAAGTAAGTTGGCCGCAGTGTTATCACTCATGGTTAT

GGCAGCACTGCATAATTCTCTTACTGTCATGCCATCCGTAAGATGCTTTTCTGTGACTGG

TGAGTACTCAACCAAGTCATTCTGAGAATAGTGTATGCGGCGACCGAGTTGCTCTTGCCC

GGCGTCAATACGGGATAATACCGCGCCACATAGCAGAACTTTAAAAGTGCTCATCATTGG

AAAACGTTCTTCGGGGCGAAAACTCTCAAGGATCTTACCGCTGTTGAGATCCAGTTCGAT

GTAACCCACTCGTGCACCCAACTGATCTTCAGCATCTTTTACTTTCACCAGCGTTTCTGG

GTGAGCAAAAACAGGAAGGCAAAATGCCGCAAAAAAGGGAATAAGGGCGACACGGAAATG

TTGAATACTCATACTCTTCCTTTTTCAATATTATTGAAGCATTTATCAGGGTTATTGTCT

CATGAGCGGATACATATTTGAATGTATTTAGAAAAATAAACAAATAGGGGTTCCGCGCAC

ATTTCCCCGAAAAGTGCCAC

Restriction sites to linearize plasmid PacI

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	GGACTAGTCAGCAAGAATAATATCAGATCAAACAG
Additional information primer 1	L6657 (SpeI); PBANKA_123880 Forward
Sequence Primer 2	CGGGATCCCTTTCTAGAGGCGTTCATAATGTG
Additional information primer 2	L6658 (BamHI); PBANKA_123880 Reverse
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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