RMgmDB - Rodent Malaria genetically modified Parasites

Back to search results

Summary

RMgm-1235

Malaria parasite	P. berghei
Genotype
Tagged	Gene model (rodent): PBANKA_1040561; Gene model (P.falciparum): Not available; Gene product: PIR protein; BIR protein (PIR7) Name tag: mCherry
Phenotype	Asexual bloodstage;

Last modified: 18 November 2016, 13:04

*RMgm-1235

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
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	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
top of page
Name of the mutant parasite
RMgm number	RMgm-1235
Principal name	2211
Alternative name
Standardized name
Is the mutant parasite cloned after genetic modification	No
top of page
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

top of page

Details of the target gene

Gene Model of Rodent Parasite PBANKA_1040561

Gene Model P. falciparum ortholog Not available

Gene product PIR protein; BIR protein

Gene product: Alternative name PIR7

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

TGGATCTCCGACAAATTAAGGGAAAAAAAAATATTGAGAAAAGCAAAGATACCTCGAGAT

ATATAGATGTTGTTAAAAATAGATTTGAATCGTATAGCTCATTTTTTAGTATTATGTTTA

CTAATGTTGGCAATAACTTATATAAAAAGTCGTTGCCAGCGCTAACAAATGTTTACGATA

AATTTATAAATTTTGCTGATAACACGATTAATTATGTGAATGGGGAGTTAAAAAAACCAA

TAAAAACTTATACATTCGATAATGGTATACCTGAGGAAAAAGGCCCAGGAGGTGACCCAT

TATCATCTCAAGAAACTCCATCTGAAGCTTCATCACCATCAAGTTCTACGGAACAAACTA

GGACATCAGAATTACCTCAGGAATCATCTAAAAAAAAAAAATATGATCAAAAGAATGAAG

AAGATTCTAAAAAAACAGTGCCAAATTTAGTGATTAAACGCGAAAATTCAGTAACCGAAG

TAACAGGAAATGGAACGACAGAAATAGATGACAATCCACTCAACTTATACAAGAAAATTG

TAATTTCAATTATAATGCTTTTAATACCCATTGTTTTAGCTATTATGTACAAGGTAAATA

AAAAGAAAACTGTGAAGTATATAATTTTAAAAATATTTCTTCACTATAAATATTATATAT

TTTTCATAATTTTATGTTAGTATTTTTCATAATTTTATGCTAGTATTTTTCATAATTTTA

TGTTAGTATTTTCCATAATTTTATGTTAGTATTTTTCATAATTTTATGTTAGTATTTTTC

ATTTGGATGGAGAAAGGAATTGAAGAAAAAAAAAAACATGAAAAAGGCTATAAATATGTT

TGGTGCAAATGAAACGACAAAAAGAGTTATAAACCCAACTGATCGAAAAAAACAAGTGCA

AATAATTATAAATTCATCTACTCAAAAAAAACAGGACAAAAAGTTTAAAAATTCATCTAC

TCAAAAAAAACAGGACAAAAAGTTTAAAAATTCATTTACTCAAAAAAAACAGGATAAAAA

GGTTACAAATTCATCTACTCAAAAAAAACAGGATAAAAAGGTTAAAAATTCCATTTATTG

GGGAAAGTATCCATTATTAAATATACACAAACTTATGAAGACCGATTCTGTACCATTTAT

TATTTTATTTTTGTTGTTTATTTTTTATGTTTATAAAAGAAAAGGCTATTATTTAGAAGG

ATCCGGTGGAGGTGGAAGATCCATGGTGAGCAAGGGCGAGGAGGATAACATGGCCATCAT

CAAGGAGTTCATGCGCTTCAAGGTGCACATGGAGGGCTCCGTGAACGGCCACGAGTTCGA

GATCGAGGGCGAGGGCGAGGGCCGCCCCTACGAGGGCACCCAGACCGCCAAGCTGAAGGT

GACCAAGGGTGGCCCCCTGCCCTTCGCCTGGGACATCCTGTCCCCTCAGTTCATGTACGG

CTCCAAGGCCTACGTGAAGCACCCCGCCGACATCCCCGACTACTTGAAGCTGTCCTTCCC

CGAGGGCTTCAAGTGGGAGCGCGTGATGAACTTCGAGGACGGCGGCGTGGTGACCGTGAC

CCAGGACTCCTCCCTGCAGGACGGCGAGTTCATCTACAAGGTGAAGCTGCGCGGCACCAA

CTTCCCCTCCGACGGCCCCGTAATGCAGAAGAAGACCATGGGCTGGGAGGCCTCCTCCGA

GCGGATGTACCCCGAGGACGGCGCCCTGAAGGGCGAGATCAAGCAGAGGCTGAAGCTGAA

GGACGGCGGCCACTACGACGCTGAGGTCAAGACCACCTACAAGGCCAAGAAGCCCGTGCA

GCTGCCCGGCGCCTACAACGTCAACATCAAGTTGGACATCACCTCCCACAACGAGGACTA

CACCATCGTGGAACAGTACGAACGCGCCGAGGGCCGCCACTCCACCGGCGGCATGGACGA

GCTGTACAAGTAATCTAGAATTATTAATATATATGAATATATATACATCGTTGTATGCGT

AATATACGTGTAACTGTTTTTTTTAATTTTTATATTAACTTTTCCTTATACATTTTAATG

CCATTAACTAATAATATTTTACTTGAAAAAATTCTAAATTTTTATAACTCTAATATAAAA

TCTGTGAATAATAAAAAATTAGCATAAAGTACGAATTAAACATCGAATATTTGTTTTAAA

GGGGCACACATTATGTGCATATAATTTTATTCTTTGAATTTAATTTTTTGCTTATAATTA

GATTAAATTAATATAATTTTTTTATTGAAAATAAAAACTTATTTTAAAAAATTAACAAAT

GGAAGTTAATGAAAAATACGAAAAAAGTAAAGGGTTAATTCTTATATGGTCGGTACCGAG

CTCGAATTCGATATCGAAATTGAAGGAAAAAACATCATTTGTGTTTATATGTTTTTTTTT

AATTTTTCAACTTTTTTTATTTAACTATGCTAATCATATTATAAACATTTTAAAAATCAA

GTCCAACTATTTATGAATCATTGAAGAGACAACATAATTATAACAGTAAAAATAATAATA

ATAATAATAATTTTGTCTTATTGTTTCAAACATATTATGTTCCATAAATATCTAAATTTT

TAATATGGTCATAAATTCATAGGATTTAATTTATATAAAATATGGCTATTCATACTAGCC

ATTTTATGTGTGTAAAGTTTTTTTTTTCTGAACAAAAAATAAAATGTTCATAATTTTAGC

TATTTACATGCATGTGCATGCACAAAAAAAAATATGCACACAACATACACATTTTTACAG

TTATAAATACAATCAATTGGTATAAATATATAAGTAAGAAAAACGGGATCAAAGCTAGCT

TCTGTATTTCCGCTAGACAGCCATCTCCATCTGGATTCGTCCGTGCGGGACGTAGCCCAC

GACCTCAAAATCCTCGGCGGTGAAATCGTCGATTTCCTTGATGCGTTCCTTGTTGAGGAT

GTTCACAATGGGGAACGGTCTCGGTTCTCTCCGCAGCTGCTCTTTTAAAGCCTCGACATG

GTTCGTGTAGACATGCGTGTTCCCCATGAAGTGAATGAACTCCTTAGGTTTTAGGTTGCA

GACGTGTGCAACCATGAGCGTCAAAAGCGAATAGGAAGCGATGTTGAAGGGGACGCCGAG

GCCGACATCGCACGACCGCTGATACATGATGCACGACAGCTCCTTCTGGTCGTTCACGTA

GAACTGGCACAACAAGTGACAAGGCGGCAGCGCCATTTCGTCCAGCGCTGCAGGATTCCA

GGCAGTCATGAGCATGCGACGATCTGTTGGATTCGTTCTCAGCATCTGGATCACATTCTT

CAGCTGGTCGACGCCCTGCCCTGTGTAGTCTGTGTGCATGTCTTTGTATGCCGCGCCGAA

GTGTCTCCACTGGAAGCCGTAGCCCGGGCCGATGTCTCCGACCTCTCGGTGGGGGAGATT

GCGCGAATCGAGGAACTCGCGTGTCACATTCTTGTCCCAGATCTTCACGCCCTTCTCAGA

AAGATGGTTTGCGTTCGTGTCGCCGCGAATGAACCACAGCAACTCTTCGAGGACCCCTTT

CCAGAACACACGCTTTGTGGTGAGAAGTGGAAAGGCCTGATCCAGCGAGTAGCGCATAGT

GCAGCCGAATTTGGAGATGACACCAACGCCCGTTCGGTCATCCATTGTCCTTCCATTGTT

AATAATGTCGGCAATGAGATCAAGGTACTGGAACTCTTCATGGCCTCTAAAGTGAACATG

CGGAACGGCCCGAATCAGTTCCTTTTGCTCGCGTTTTTTCCGGTCTTCTTCGTCCATCCA

CGCCAACACCGGGGCAATGGCTGCGGCCGAAGAAGGAGCCTGCAACCCGTGCACGGGAGT

TGTCTCCCTCGTGGACGTCAAGGAGCTCATTGCGTTGCTCGGTTCCGCAGTGGCTGCGTC

GTCAGTCTTCCTTCTCTTCTCGAGAACCACAAAGTCGTAGGGAACCCCGTTGTCTGAGAA

GGTCTTGGAAATGAAGATGGGTCGATACGTCGCTTCATTGTCCTTCTCTCTTCCGAGCTC

CGGACAAAAGGGAACGAACACAGACTCGGCAGGAGCTGCAGCCTGCGCAGCAGTTGATTT

GTTTGAAAGAATGTCATCTCCGGGGAACGCAGGGAAGAAAACGTCGCACGGAAACTCGCG

GGCTACACGCGTGATGTACAGGTGAGAGGCAACGCCCAGAGACAGCGCTGCCTCGTACAG

TCCCGCTCCTCCCACGACAAAAATCTGGTCGACAGAATCCTTGTACTCTTCCTCCAGAAG

GCTGAGAGCTGCTGGGAGTGAAGCACAGACTCGGACGCGCTGCTGGCCTTCAGCTTGAGG

CTTCTCCGCCGCAATGTCTTCTTCTTTGAGGGAAGAGGAAACGACGATGTTCAATCTGTC

CACGAGGGGTCTAAACTTTCGAGGCATGCTTTCCCAGGTTTTCCGTCCCATGACAACGGC

GTTGAATCTCTTGCCGACTGATGGAGAGGGAAGTCCAGAGTCGCCCGTCTTTGCAAATTT

CCTGGGAAGCCACCCGTTCAGGCGACTGGCTTCTTCGGGCGTCGTTTTTGTCACACGAGA

AAAGTGTTTGAAATCTGTGGTCAAGTGGGGCCACGGGAGGCCGTTGTTGATGCCGATGCC

CCTCTTGGGGGTCATCGCGACGACCAGACACACCGGTTTATGCATTGATCCTTTGTAACA

TTTAGGTGTGTATTTATATATATAAGCAATATACAAATGAAAATACTATTATAAAACGGA

AACTCAAAAAATGTTGAAACAAATAGTCAATTTTTAAAATGAATGTGTAAATTGTTTAAG

TTTTATGACAAAAAAAGGAAAAGTGTATATAAAATAAAAATTGGAAAAAATTTATATATT

ACATGCCATATAATACCATTTATTTACTATTTATATCTTAATATAAAAAATTGTAGCAAA

AATATATAATCGTATTTTATATATTTAATATTATATAACCCCTTAAAAAAAAAAATTTAA

TTGAAAAAATTTTAATTTATGCACTTTTTTATTGTATTATATATATTCCGTTATTTTAAT

TACACATAAACTTAATAGAAAAATTAATTCGCTGCTAAATAAAAATTATAGGGCTATATA

TGATGGGATCATATTCTATCTTTCTCTTTTTCAAATYCTTTATATTTTAGAATACAAATC

GCTTTAAACAATAAAAAAGAGGTATATCCTACCCGAATTGACACACACAATAAATAATAA

ATATATAAATAATAAAAATATAAATATATAAATTATAAATATATAAATTATATATATAAA

TATATTAACATGTGTGTAATTTGTTAAAATTGATATTAAACAGCTTTGCATGGGGAAAGA

GTGGCTTTTATAAGCGAATCATATAATACAGTGTTAATTTTAATATATAAAATATAATTT

CCTTAAATTGTTATTTAACATTTTTAATTGCCTACCCARGGTAATATAATATCCAAATGA

CAATCAAAGATAAATGAAATACCGCTCCATTTTTCCTATATATCCATTTTCATTTTGTGT

AAATGTTCATAAAATTTTATCATATACTCATAAAAAACAGGGGAAAAATGAAAAAATGAA

CGTCCACCCCGTGTGAATATGCTCATTTTGTATACTATACATTTTTTTTTATATAATTTA

TATTTTCCAAATAAAAAAAATGTTTGTACATGTATATATCAATATTGTGTTATGGCATGA

TGAAAAACAATTATTTTATTCGTTTTTAAATTTGTAGAATTTCCCTAAATTATTAATGAT

TTCATATTATAATATATGTATCTATATATTTTTAATGTGTAATTCAAAATGGCGAAAAAG

AAACAAACCATACTTTATAAGCAACTCTCTTTGAGCACTTACCATAAAAAATTATACTAC

TAAATTCAAATTAAATAAAAAAATATATATTATAAAATTGGCTTTAATTATTTATAAGCA

CTTGCTTTGTTTTACTCAAAAAGTATTGGCATAAGTATAAACATAAAACCAATTTTTCAC

AAAATATATTATATGTCCATACAACTATATCCGAACYTTCAAAACACATCCAAATTTCCA

TTTACCACATGCAAGCAAAAGGATATTAAATAAAAAAAAATTATTTATATTTTCATGATC

ACTTAAATAATATTTAAAAAAAAAAGAGACCATATATAAAATGTATAATTTAAATATAGC

TAACTTATTAAAAAAAATGAATATTGAGCATATATCCATAATTTAAAACACTTCTATTTT

ATGTTATTTGATATATAAGTCGATAAATTATTGCATGTATGCATATAGACAGATTAATAT

ACACCAAAATAATAAAAATTATATATAGTTCATATATTATTCTGAATAATAAATTAGCTT

TCCTCTTAATTCACTTGGTATGTATATATATATATATATATATATATATATATATATGGA

TAAATAAATATAAGGTCTATAACAGGAAAAAATGTGTATTAACTTTTTTTTATAATTATT

TTATATTTTTTCTCCATTTATTTAAATATGTTAAAATTATTTCCCATTTTATAAACAAAA

ATTAATCCATAAAATAGCAAAATAACAAAATGACAAAATAACAAAAACAAAAAAACGCAT

TATATGAGTTCATTTTACACAATCCAAAGAAAAAGAAAATATGCTTAAAATATTCATAAC

ACACTTTTAAGCTATAAAAATATATATATTTATTTGTGTCTATATTACCAACTCAATTTA

ATAGATGTGTTATGTGATTAATTCATACACAAACATACAAAAATAAACACCATTTTTGAA

AAGATTAATTTGAAATCATATATGTTATTATTATATATTTTTATATCAATGAAATTTAAA

AAAATAAAAGGGAAATCAATGTATTAAAAATAATTATATGTTATTTTATTTCCACAATAT

TTATTTATTATTTATTGTTGACCTGCAGGCATGCAAGCTAGCTTATCGATACCGTCGACC

TCGAGGGGGGGCCCGGTACCCAGCTTTTGTTCCCTTTAGTGAGGGTTAATTGCGCGCTTG

GCGTAATCATGGTCATAGCTGTTTCCTGTGTGAAATTGTTATCCGCTCACAATTCCACAC

AACATACGAGCCGGAAGCATAAAGTGTAAAGCCTGGGGTGCCTAATGAGTGAGCTAACTC

ACATTAATTGCGTTGCGCTCACTGCCCGCTTTCCAGTCGGGAAACCTGTCGTGCCAGCTG

CATTAATGAATCGGCCAACGCGCGGGGAGAGGCGGTTTGCGTATTGGGCGCTCTTCCGCT

TCCTCGCTCACTGACTCGCTGCGCTCGGTCGTTCGGCTGCGGCGAGCGGTATCAGCTCAC

TCAAAGGCGGTAATACGGTTATCCACAGAATCAGGGGATAACGCAGGAAAGAACATGTGA

GCAAAAGGCCAGCAAAAGGCCAGGAACCGTAAAAAGGCCGCGTTGCTGGCGTTTTTCCAT

AGGCTCCGCCCCCCTGACGAGCATCACAAAAATCGACGCTCAAGTCAGAGGTGGCGAAAC

CCGACAGGACTATAAAGATACCAGGCGTTTCCCCCTGGAAGCTCCCTCGTGCGCTCTCCT

GTTCCGACCCTGCCGCTTACCGGATACCTGTCCGCCTTTCTCCCTTCGGGAAGCGTGGCG

CTTTCTCATAGCTCACGCTGTAGGTATCTCAGTTCGGTGTAGGTCGTTCGCTCCAAGCTG

GGCTGTGTGCACGAACCCCCCGTTCAGCCCGACCGCTGCGCCTTATCCGGTAACTATCGT

CTTGAGTCCAACCCGGTAAGACACGACTTATCGCCACTGGCAGCAGCCACTGGTAACAGG

ATTAGCAGAGCGAGGTATGTAGGCGGTGCTACAGAGTTCTTGAAGTGGTGGCCTAACTAC

GGCTACACTAGAAGGACAGTATTTGGTATCTGCGCTCTGCTGAAGCCAGTTACCTTCGGA

AAAAGAGTTGGTAGCTCTTGATCCGGCAAACAAACCACCGCTGGTAGCGGTGGTTTTTTT

GTTTGCAAGCAGCAGATTACGCGCAGAAAAAAAGGATCTCAAGAAGATCCTTTGATCTTT

TCTACGGGGTCTGACGCTCAGTGGAACGAAAACTCACGTTAAGGGATTTTGGTCATGAGA

TTATCAAAAAGGATCTTCACCTAGATCCTTTTAAATTAAAAATGAAGTTTTAAATCAATC

TAAAGTATATATGAGTAAACTTGGTCTGACAGTTACCAATGCTTAATCAGTGAGGCACCT

ATCTCAGCGATCTGTCTATTTCGTTCATCCATAGTTGCCTGACTCCCCGTCGTGTAGATA

ACTACGATACGGGAGGGCTTACCATCTGGCCCCAGTGCTGCAATGATACCGCGAGACCCA

CGCTCACCGGCTCCAGATTTATCAGCAATAAACCAGCCAGCCGGAAGGGCCGAGCGCAGA

AGTGGTCCTGCAACTTTATCCGCCTCCATCCAGTCTATTAATTGTTGCCGGGAAGCTAGA

GTAAGTAGTTCGCCAGTTAATAGTTTGCGCAACGTTGTTGCCATTGCTACAGGCATCGTG

GTGTCACGCTCGTCGTTTGGTATGGCTTCATTCAGCTCCGGTTCCCAACGATCAAGGCGA

GTTACATGATCCCCCATGTTGTGCAAAAAAGCGGTTAGCTCCTTCGGTCCTCCGATCGTT

GTCAGAAGTAAGTTGGCCGCAGTGTTATCACTCATGGTTATGGCAGCACTGCATAATTCT

CTTACTGTCATGCCATCCGTAAGATGCTTTTCTGTGACTGGTGAGTACTCAACCAAGTCA

TTCTGAGAATAGTGTATGCGGCGACCGAGTTGCTCTTGCCCGGCGTCAATACGGGATAAT

ACCGCGCCACATAGCAGAACTTTAAAAGTGCTCATCATTGGAAAACGTTCTTCGGGGCGA

AAACTCTCAAGGATCTTACCGCTGTTGAGATCCAGTTCGATGTAACCCACTCGTGCACCC

AACTGATCTTCAGCATCTTTTACTTTCACCAGCGTTTCTGGGTGAGCAAAAACAGGAAGG

CAAAATGCCGCAAAAAAGGGAATAAGGGCGACACGGAAATGTTGAATACTCATACTCTTC

CTTTTTCAATATTATTGAAGCATTTATCAGGGTTATTGTCTCATGAGCGGATACATATTT

GAATGTATTTAGAAAAATAAACAAATAGGGGTTCCGCGCACATTTCCCCGAAAAGTGCCA

CCTAAATTGTAAGCGTTAATATTTTGTTAAAATTCGCGTTAAATTTTTGTTAAATCAGCT

CATTTTTTAACCAATAGGCCGAAATCGGCAAAATCCCTTATAAATCAAAAGAATAGACCG

AGATAGGGTTGAGTGTTGTTCCAGTTTGGAACAAGAGTCCACTATTAAAGAACGTGGACT

CCAACGTCAAAGGGCGAAAAACCGTCTATCAGGGCGATGGCCCACTACGTGAACCATCAC

CCTAATCAAGTTTTTTGGGGTCGAGGTGCCGTAAAGCACTAAATCGGAACCCTAAAGGGA

GCCCCCGATTTAGAGCTTGACGGGGAAAGCCGGCGAACGTGGCGAGAAAGGAAGGGAAGA

AAGCGAAAGGAGCGGGCGCTAGGGCGCTGGCAAGTGTAGCGGTCACGCTGCGCGTAACCA

CCACACCCGCCGCGCTTAATGCGCCGCTACAGGGCGCGTCCCATTCGCCATTCAGGCTGC

GCAACTGTTGGGAAGGGCGATCGGTGCGGGCCTCTTCGCTATTACGCCAGCTGGCGAAAG

GGGGATGTGCTGCAAGGCGATTAAGTTGGGTAACGCCAGGGTTTTCCCAGTCACGACGTT

GTAAAACGACGGCCAGTGAGCGCGCGTAATACGACTCACTATAGGGCGAATTGGAGCTCC

ACCGCGGTGGCGGCCGCTCTAGAACTAGT

Restriction sites to linearize plasmid HindIII

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

top of page