RMgmDB - Rodent Malaria genetically modified Parasites

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Summary

RMgm-710

Malaria parasite	P. berghei
Genotype
Tagged	Gene model (rodent): PBANKA_1137800; Gene model (P.falciparum): PF3D7_1361800; Gene product: glideosome-associated connector Name tag: mCherry
Phenotype	Asexual bloodstage;

Last modified: 2 December 2012, 15:40

*RMgm-710

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-710
Principal name	1919
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 RMgm-725: mutant lacking expression of PBANKA_113780

Tagged: Mutant parasite with a tagged gene

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Details of the target gene

Gene Model of Rodent Parasite PBANKA_1137800

Gene Model P. falciparum ortholog PF3D7_1361800

Gene product glideosome-associated connector

Gene product: Alternative name

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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

CCGCGGTGGCGGCCGCTCTAGAACTAGTTGTGGTGGAATACCTACTATAGCAAATTTGAT

ATCTAAATCTGTTAAAAAGTCAATTGTAGTTTTAGAAGCATTACGTATATTATTATGTGT

ATTATATTATACAAAAAATGTCGAAGGTGTAGTTAATGAATATTCTGAAGAAGACGAAGA

ATTATTCAATGCTAGATTAGGGGGATGGTATAATATAAGTATGGACAAAGAAATGATAGA

TGTAATATTACAATCTGTTTTAACATGTTCAAATGATGAGAATCATCAAAAACAATTGAG

ATTACAAAAAGTGTCTCTTGGTTTATTAGCTTATTTTGCATATCATAGGTTAGGTATTAT

TTCTATGACTGCATCTGGTTTTGATATTTTGGCTAAAAACAATTTAAATCATTTCGGTGG

TGATATGGTAATTATGCAATTGTTGGCTATATGTATAGATAACATTGCGATGAATTCAGC

AGAAATATACGATATGACAATTACGAGAGATATTATAAAATTATTTAAATCATCTATATC

TAAAATCCCAAATAAAAAAGATAATAAGCAAATTGTACAAAGTATTGAAAAAACAGTAGA

AGCAATGGGTTCAGATGGAGATCCTTTAGATGCATTTAAAGATACTATATTAACTTTTGA

TTTTAGTTTATCAGAATTTGATAAAGATCCATATGTTAATGGTGTACACGATTTACCTCA

AAATATCAAAGAAGCGTTGAGAACAGGTGGTCAATATAAAATATACCATAAAAGTGATAA

AAGAACATTGTTCAAATGGAAAGCTAGTCAAGATTTAGGGACCTTAGAATGGACAGTAGG

AGAAAATGTAGATCGTATATTTAAAATATCAGTAGTACGTATTAAAAATATATCCAAAGG

ATTAGTACATCCTTTGTTAAAAGCGGCTAATAAATATGAGCCTAGAAAAGTCAATTCAAA

AGTAGTATTATGCGTTTATGGACCCCCTACTGAAGATTTTCCAGAGGGCCTAGAGTTGCC

AATTAAAACAAAAACAAACAAAGAAAGAGATGCATTCGCCGATCTATTAATTCTTTGGCG

TGATGCTGCTAGTTATAACTATGGATCCGGTGGAGGTGGAAGATCCATGGTGAGCAAGGG

CGAGGAGGATAACATGGCCATCATCAAGGAGTTCATGCGCTTCAAGGTGCACATGGAGGG

CTCCGTGAACGGCCACGAGTTCGAGATCGAGGGCGAGGGCGAGGGCCGCCCCTACGAGGG

CACCCAGACCGCCAAGCTGAAGGTGACCAAGGGTGGCCCCCTGCCCTTCGCCTGGGACAT

CCTGTCCCCTCAGTTCATGTACGGCTCCAAGGCCTACGTGAAGCACCCCGCCGACATCCC

CGACTACTTGAAGCTGTCCTTCCCCGAGGGCTTCAAGTGGGAGCGCGTGATGAACTTCGA

GGACGGCGGCGTGGTGACCGTGACCCAGGACTCCTCCCTGCAGGACGGCGAGTTCATCTA

CAAGGTGAAGCTGCGCGGCACCAACTTCCCCTCCGACGGCCCCGTAATGCAGAAGAAGAC

CATGGGCTGGGAGGCCTCCTCCGAGCGGATGTACCCCGAGGACGGCGCCCTGAAGGGCGA

GATCAAGCAGAGGCTGAAGCTGAAGGACGGCGGCCACTACGACGCTGAGGTCAAGACCAC

CTACAAGGCCAAGAAGCCCGTGCAGCTGCCCGGCGCCTACAACGTCAACATCAAGTTGGA

CATCACCTCCCACAACGAGGACTACACCATCGTGGAACAGTACGAACGCGCCGAGGGCCG

CCACTCCACCGGCGGCATGGACGAGCTGTACAAGTAATCTAGAATTATTAATATATATGA

ATATATATACATCGTTGTATGCGTAATATACGTGTAACTGTTTTTTTTAATTTTTATATT

AACTTTTCCTTATACATTTTAATGCCATTAACTAATAATATTTTACTTGAAAAAATTCTA

AATTTTTATAACTCTAATATAAAATCTGTGAATAATAAAAAATTAGCATAAAGTACGAAT

TAAACATCGAATATTTGTTTTAAAGGGGCACACATTATGTGCATATAATTTTATTCTTTG

AATTTAATTTTTTGCTTATAATTAGATTAAATTAATATAATTTTTTTATTGAAAATAAAA

ACTTATTTTAAAAAATTAACAAATGGAAGTTAATGAAAAATACGAAAAAAGTAAAGGGTT

AATTCTTATATGGTCGGTACCGAGCTCGAATTCGATATCGAAATTGAAGGAAAAAACATC

ATTTGTGTTTATATGTTTTTTTTTAATTTTTCAACTTTTTTTATTTAACTATGCTAATCA

TATTATAAACATTTTAAAAATCAAGTCCAACTATTTATGAATCATTGAAGAGACAACATA

ATTATAACAGTAAAAATAATAATAATAATAATAATTTTGTCTTATTGTTTCAAACATATT

ATGTTCCATAAATATCTAAATTTTTAATATGGTCATAAATTCATAGGATTTAATTTATAT

AAAATATGGCTATTCATACTAGCCATTTTATGTGTGTAAAGTTTTTTTTTTCTGAACAAA

AAATAAAATGTTCATAATTTTAGCTATTTACATGCATGTGCATGCACAAAAAAAAATATG

CACACAACATACACATTTTTACAGTTATAAATACAATCAATTGGTATAAATATATAAGTA

AGAAAAACGGGATCAAAGCTAGCTTCTGTATTTCCGCTAGACAGCCATCTCCATCTGGAT

TCGTCCGTGCGGGACGTAGCCCACGACCTCAAAATCCTCGGCGGTGAAATCGTCGATTTC

CTTGATGCGTTCCTTGTTGAGGATGTTCACAATGGGGAACGGTCTCGGTTCTCTCCGCAG

CTGCTCTTTTAAAGCCTCGACATGGTTCGTGTAGACATGCGTGTTCCCCATGAAGTGAAT

GAACTCCTTAGGTTTTAGGTTGCAGACGTGTGCAACCATGAGCGTCAAAAGCGAATAGGA

AGCGATGTTGAAGGGGACGCCGAGGCCGACATCGCACGACCGCTGATACATGATGCACGA

CAGCTCCTTCTGGTCGTTCACGTAGAACTGGCACAACAAGTGACAAGGCGGCAGCGCCAT

TTCGTCCAGCGCTGCAGGATTCCAGGCAGTCATGAGCATGCGACGATCTGTTGGATTCGT

TCTCAGCATCTGGATCACATTCTTCAGCTGGTCGACGCCCTGCCCTGTGTAGTCTGTGTG

CATGTCTTTGTATGCCGCGCCGAAGTGTCTCCACTGGAAGCCGTAGCCCGGGCCGATGTC

TCCGACCTCTCGGTGGGGGAGATTGCGCGAATCGAGGAACTCGCGTGTCACATTCTTGTC

CCAGATCTTCACGCCCTTCTCAGAAAGATGGTTTGCGTTCGTGTCGCCGCGAATGAACCA

CAGCAACTCTTCGAGGACCCCTTTCCAGAACACACGCTTTGTGGTGAGAAGTGGAAAGGC

CTGATCCAGCGAGTAGCGCATAGTGCAGCCGAATTTGGAGATGACACCAACGCCCGTTCG

GTCATCCATTGTCCTTCCATTGTTAATAATGTCGGCAATGAGATCAAGGTACTGGAACTC

TTCATGGCCTCTAAAGTGAACATGCGGAACGGCCCGAATCAGTTCCTTTTGCTCGCGTTT

TTTCCGGTCTTCTTCGTCCATCCACGCCAACACCGGGGCAATGGCTGCGGCCGAAGAAGG

AGCCTGCAACCCGTGCACGGGAGTTGTCTCCCTCGTGGACGTCAAGGAGCTCATTGCGTT

GCTCGGTTCCGCAGTGGCTGCGTCGTCAGTCTTCCTTCTCTTCTCGAGAACCACAAAGTC

GTAGGGAACCCCGTTGTCTGAGAAGGTCTTGGAAATGAAGATGGGTCGATACGTCGCTTC

ATTGTCCTTCTCTCTTCCGAGCTCCGGACAAAAGGGAACGAACACAGACTCGGCAGGAGC

TGCAGCCTGCGCAGCAGTTGATTTGTTTGAAAGAATGTCATCTCCGGGGAACGCAGGGAA

GAAAACGTCGCACGGAAACTCGCGGGCTACACGCGTGATGTACAGGTGAGAGGCAACGCC

CAGAGACAGCGCTGCCTCGTACAGTCCCGCTCCTCCCACGACAAAAATCTGGTCGACAGA

ATCCTTGTACTCTTCCTCCAGAAGGCTGAGAGCTGCTGGGAGTGAAGCACAGACTCGGAC

GCGCTGCTGGCCTTCAGCTTGAGGCTTCTCCGCCGCAATGTCTTCTTCTTTGAGGGAAGA

GGAAACGACGATGTTCAATCTGTCCACGAGGGGTCTAAACTTTCGAGGCATGCTTTCCCA

GGTTTTCCGTCCCATGACAACGGCGTTGAATCTCTTGCCGACTGATGGAGAGGGAAGTCC

AGAGTCGCCCGTCTTTGCAAATTTCCTGGGAAGCCACCCGTTCAGGCGACTGGCTTCTTC

GGGCGTCGTTTTTGTCACACGAGAAAAGTGTTTGAAATCTGTGGTCAAGTGGGGCCACGG

GAGGCCGTTGTTGATGCCGATGCCCCTCTTGGGGGTCATCGCGACGACCAGACACACCGG

TTTATGCATTGATCCTTTGTAACATTTAGGTGTGTATTTATATATATAAGCAATATACAA

ATGAAAATACTATTATAAAACGGAAACTCAAAAAATGTTGAAACAAATAGTCAATTTTTA

AAATGAATGTGTAAATTGTTTAAGTTTTATGACAAAAAAAGGAAAAGTGTATATAAAATA

AAAATTGGAAAAAATTTATATATTACATGCCATATAATACCATTTATTTACTATTTATAT

CTTAATATAAAAAATTGTAGCAAAAATATATAATCGTATTTTATATATTTAATATTATAT

AACCCCTTAAAAAAAAAAATTTAATTGAAAAAATTTTAATTTATGCACTTTTTTATTGTA

TTATATATATTCCGTTATTTTAATTACACATAAACTTAATAGAAAAATTAATTCGCTGCT

AAATAAAAATTATAGGGCTATATATGATGGGATCATATTCTATCTTTCTCTTTTTCAAAT

YCTTTATATTTTAGAATACAAATCGCTTTAAACAATAAAAAAGAGGTATATCCTACCCGA

ATTGACACACACAATAAATAATAAATATATAAATAATAAAAATATAAATATATAAATTAT

AAATATATAAATTATATATATAAATATATTAACATGTGTGTAATTTGTTAAAATTGATAT

TAAACAGCTTTGCATGGGGAAAGAGTGGCTTTTATAAGCGAATCATATAATACAGTGTTA

ATTTTAATATATAAAATATAATTTCCTTAAATTGTTATTTAACATTTTTAATTGCCTACC

CARGGTAATATAATATCCAAATGACAATCAAAGATAAATGAAATACCGCTCCATTTTTCC

TATATATCCATTTTCATTTTGTGTAAATGTTCATAAAATTTTATCATATACTCATAAAAA

ACAGGGGAAAAATGAAAAAATGAACGTCCACCCCGTGTGAATATGCTCATTTTGTATACT

ATACATTTTTTTTTATATAATTTATATTTTCCAAATAAAAAAAATGTTTGTACATGTATA

TATCAATATTGTGTTATGGCATGATGAAAAACAATTATTTTATTCGTTTTTAAATTTGTA

GAATTTCCCTAAATTATTAATGATTTCATATTATAATATATGTATCTATATATTTTTAAT

GTGTAATTCAAAATGGCGAAAAAGAAACAAACCATACTTTATAAGCAACTCTCTTTGAGC

ACTTACCATAAAAAATTATACTACTAAATTCAAATTAAATAAAAAAATATATATTATAAA

ATTGGCTTTAATTATTTATAAGCACTTGCTTTGTTTTACTCAAAAAGTATTGGCATAAGT

ATAAACATAAAACCAATTTTTCACAAAATATATTATATGTCCATACAACTATATCCGAAC

YTTCAAAACACATCCAAATTTCCATTTACCACATGCAAGCAAAAGGATATTAAATAAAAA

AAAATTATTTATATTTTCATGATCACTTAAATAATATTTAAAAAAAAAAGAGACCATATA

TAAAATGTATAATTTAAATATAGCTAACTTATTAAAAAAAATGAATATTGAGCATATATC

CATAATTTAAAACACTTCTATTTTATGTTATTTGATATATAAGTCGATAAATTATTGCAT

GTATGCATATAGACAGATTAATATACACCAAAATAATAAAAATTATATATAGTTCATATA

TTATTCTGAATAATAAATTAGCTTTCCTCTTAATTCACTTGGTATGTATATATATATATA

TATATATATATATATATATATGGATAAATAAATATAAGGTCTATAACAGGAAAAAATGTG

TATTAACTTTTTTTTATAATTATTTTATATTTTTTCTCCATTTATTTAAATATGTTAAAA

TTATTTCCCATTTTATAAACAAAAATTAATCCATAAAATAGCAAAATAACAAAATGACAA

AATAACAAAAACAAAAAAACGCATTATATGAGTTCATTTTACACAATCCAAAGAAAAAGA

AAATATGCTTAAAATATTCATAACACACTTTTAAGCTATAAAAATATATATATTTATTTG

TGTCTATATTACCAACTCAATTTAATAGATGTGTTATGTGATTAATTCATACACAAACAT

ACAAAAATAAACACCATTTTTGAAAAGATTAATTTGAAATCATATATGTTATTATTATAT

ATTTTTATATCAATGAAATTTAAAAAAATAAAAGGGAAATCAATGTATTAAAAATAATTA

TATGTTATTTTATTTCCACAATATTTATTTATTATTTATTGTTGACCTGCAGGCATGCAA

GCTTATCGATACCGTCGACCTCGAGGGGGGGCCCGGTACCCAGCTTTTGTTCCCTTTAGT

GAGGGTTAATTGCGCGCTTGGCGTAATCATGGTCATAGCTGTTTCCTGTGTGAAATTGTT

ATCCGCTCACAATTCCACACAACATACGAGCCGGAAGCATAAAGTGTAAAGCCTGGGGTG

CCTAATGAGTGAGCTAACTCACATTAATTGCGTTGCGCTCACTGCCCGCTTTCCAGTCGG

GAAACCTGTCGTGCCAGCTGCATTAATGAATCGGCCAACGCGCGGGGAGAGGCGGTTTGC

GTATTGGGCGCTCTTCCGCTTCCTCGCTCACTGACTCGCTGCGCTCGGTCGTTCGGCTGC

GGCGAGCGGTATCAGCTCACTCAAAGGCGGTAATACGGTTATCCACAGAATCAGGGGATA

ACGCAGGAAAGAACATGTGAGCAAAAGGCCAGCAAAAGGCCAGGAACCGTAAAAAGGCCG

CGTTGCTGGCGTTTTTCCATAGGCTCCGCCCCCCTGACGAGCATCACAAAAATCGACGCT

CAAGTCAGAGGTGGCGAAACCCGACAGGACTATAAAGATACCAGGCGTTTCCCCCTGGAA

GCTCCCTCGTGCGCTCTCCTGTTCCGACCCTGCCGCTTACCGGATACCTGTCCGCCTTTC

TCCCTTCGGGAAGCGTGGCGCTTTCTCATAGCTCACGCTGTAGGTATCTCAGTTCGGTGT

AGGTCGTTCGCTCCAAGCTGGGCTGTGTGCACGAACCCCCCGTTCAGCCCGACCGCTGCG

CCTTATCCGGTAACTATCGTCTTGAGTCCAACCCGGTAAGACACGACTTATCGCCACTGG

CAGCAGCCACTGGTAACAGGATTAGCAGAGCGAGGTATGTAGGCGGTGCTACAGAGTTCT

TGAAGTGGTGGCCTAACTACGGCTACACTAGAAGGACAGTATTTGGTATCTGCGCTCTGC

TGAAGCCAGTTACCTTCGGAAAAAGAGTTGGTAGCTCTTGATCCGGCAAACAAACCACCG

CTGGTAGCGGTGGTTTTTTTGTTTGCAAGCAGCAGATTACGCGCAGAAAAAAAGGATCTC

AAGAAGATCCTTTGATCTTTTCTACGGGGTCTGACGCTCAGTGGAACGAAAACTCACGTT

AAGGGATTTTGGTCATGAGATTATCAAAAAGGATCTTCACCTAGATCCTTTTAAATTAAA

AATGAAGTTTTAAATCAATCTAAAGTATATATGAGTAAACTTGGTCTGACAGTTACCAAT

GCTTAATCAGTGAGGCACCTATCTCAGCGATCTGTCTATTTCGTTCATCCATAGTTGCCT

GACTCCCCGTCGTGTAGATAACTACGATACGGGAGGGCTTACCATCTGGCCCCAGTGCTG

CAATGATACCGCGAGACCCACGCTCACCGGCTCCAGATTTATCAGCAATAAACCAGCCAG

CCGGAAGGGCCGAGCGCAGAAGTGGTCCTGCAACTTTATCCGCCTCCATCCAGTCTATTA

ATTGTTGCCGGGAAGCTAGAGTAAGTAGTTCGCCAGTTAATAGTTTGCGCAACGTTGTTG

CCATTGCTACAGGCATCGTGGTGTCACGCTCGTCGTTTGGTATGGCTTCATTCAGCTCCG

GTTCCCAACGATCAAGGCGAGTTACATGATCCCCCATGTTGTGCAAAAAAGCGGTTAGCT

CCTTCGGTCCTCCGATCGTTGTCAGAAGTAAGTTGGCCGCAGTGTTATCACTCATGGTTA

TGGCAGCACTGCATAATTCTCTTACTGTCATGCCATCCGTAAGATGCTTTTCTGTGACTG

GTGAGTACTCAACCAAGTCATTCTGAGAATAGTGTATGCGGCGACCGAGTTGCTCTTGCC

CGGCGTCAATACGGGATAATACCGCGCCACATAGCAGAACTTTAAAAGTGCTCATCATTG

GAAAACGTTCTTCGGGGCGAAAACTCTCAAGGATCTTACCGCTGTTGAGATCCAGTTCGA

TGTAACCCACTCGTGCACCCAACTGATCTTCAGCATCTTTTACTTTCACCAGCGTTTCTG

GGTGAGCAAAAACAGGAAGGCAAAATGCCGCAAAAAAGGGAATAAGGGCGACACGGAAAT

GTTGAATACTCATACTCTTCCTTTTTCAATATTATTGAAGCATTTATCAGGGTTATTGTC

TCATGAGCGGATACATATTTGAATGTATTTAGAAAAATAAACAAATAGGGGTTCCGCGCA

CATTTCCCCGAAAAGTGCCAC

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

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	AACTAGTTGTGGTGGAATACCTACTATAGC
Additional information primer 1	L6455 (SpeI); PBANKA_113780 Forward
Sequence Primer 2	CGGGATCCATAGTTATAACTAGCAGCATCAC
Additional information primer 2	L6456 (BamHI); PBANKA_113780 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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