RMgmDB - Rodent Malaria genetically modified Parasites

Summary

RMgm-782
Malaria parasiteP. berghei
Genotype
TaggedGene model (rodent): PBANKA_0836400; Gene model (P.falciparum): Not available; Gene product: Plasmodium exported protein, unknown function
Name tag: mCherry
Phenotype Asexual bloodstage;
Last modified: 11 August 2014, 19:05
  *RMgm-782
Successful modificationThe parasite was generated by the genetic modification
The mutant contains the following genetic modification(s) Gene tagging
Reference (PubMed-PMID number) Not published (yet)
MR4 number
Parent parasite used to introduce the genetic modification
Rodent Malaria ParasiteP. berghei
Parent strain/lineP. berghei ANKA
Name parent line/clone P. berghei ANKA cl15cy1
Other information parent lineA reference wild type clone from the ANKA strain of P. berghei (PubMed: PMID: 17406255)
The mutant parasite was generated by
Name PI/ResearcherJ.A. Eken; O. Klop; B.M.D. Franke-Fayard; C.J. Janse
Name Group/DepartmentLeiden Malaria Research Group
Name InstituteLeiden University Medical Center (LUMC)
CityLeiden
CountryThe Netherlands
Name of the mutant parasite
RMgm numberRMgm-782
Principal name2192
Alternative name
Standardized name
Is the mutant parasite cloned after genetic modificationNo
Phenotype
Asexual blood stageExpression of the mCherry-tagged protein in blood stages
Gametocyte/GameteNot tested
Fertilization and ookineteNot tested
OocystNot tested
SporozoiteNot tested
Liver stageNot tested
Additional remarks phenotype

Mutant/mutation
The mutant expresses a C-terminal mCherry-tagged version of  PBANKA_083640.
The mutant is not cloned and genotype analysis indicates the presence of episomes (see Figure below). We included this mutant in the database since it shows a clear export into the cytoplasm of the host red blood cell (see Figure below).

Protein (function)

Phenotype
The protein is expressed in blood stages. Evidence for transport into the host RBC (see Figure below).
The phenotype and expression have not been analysed in detail.

Additional information
 


Figure: mCherry::PBANKA_PBANKA_0836400 expression in blood stages and export into the cytoplasm of the host red blood cell. Nuclear staining with Hoechst (blue); erthrocyte membrane staining with anti-TER antibodies (green). Southern analysis of separated chromosomes (right) confirms integration of the construct into chromosome 8 (hybridisation with the 3'UTR Pbdhfr/ts gene recognizing the integrated construct in chr. 8 and the endogenous dhfr/ts gene on chr. 7); however, the (strong) additional hybridisation indicates also the presence of episomes/plasmids.

Other mutants


  Tagged: Mutant parasite with a tagged gene
Details of the target gene
Gene Model of Rodent Parasite PBANKA_0836400
Gene Model P. falciparum ortholog Not available
Gene productPlasmodium exported protein, unknown function
Gene product: Alternative name
Details of the genetic modification
Name of the tagmCherry
Details of taggingC-terminal
Additional remarks: tagging
Commercial source of tag-antibodies
Type of plasmid/construct(Linear) plasmid single cross-over
PlasmoGEM (Sanger) construct/vector usedNo
Modified PlasmoGEM construct/vector usedNo
Plasmid/construct map
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Plasmid/construct sequence
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TATCGATACCGTCGACCTCGAGGGGGGGCCCGGTACCCAGCTTTTGTTCCCTTTAGTGAG
GGTTAATTGCGCGCTTGGCGTAATCATGGTCATAGCTGTTTCCTGTGTGAAATTGTTATC
CGCTCACAATTCCACACAACATACGAGCCGGAAGCATAAAGTGTAAAGCCTGGGGTGCCT
AATGAGTGAGCTAACTCACATTAATTGCGTTGCGCTCACTGCCCGCTTTCCAGTCGGGAA
ACCTGTCGTGCCAGCTGCATTAATGAATCGGCCAACGCGCGGGGAGAGGCGGTTTGCGTA
TTGGGCGCTCTTCCGCTTCCTCGCTCACTGACTCGCTGCGCTCGGTCGTTCGGCTGCGGC
GAGCGGTATCAGCTCACTCAAAGGCGGTAATACGGTTATCCACAGAATCAGGGGATAACG
CAGGAAAGAACATGTGAGCAAAAGGCCAGCAAAAGGCCAGGAACCGTAAAAAGGCCGCGT
TGCTGGCGTTTTTCCATAGGCTCCGCCCCCCTGACGAGCATCACAAAAATCGACGCTCAA
GTCAGAGGTGGCGAAACCCGACAGGACTATAAAGATACCAGGCGTTTCCCCCTGGAAGCT
CCCTCGTGCGCTCTCCTGTTCCGACCCTGCCGCTTACCGGATACCTGTCCGCCTTTCTCC
CTTCGGGAAGCGTGGCGCTTTCTCATAGCTCACGCTGTAGGTATCTCAGTTCGGTGTAGG
TCGTTCGCTCCAAGCTGGGCTGTGTGCACGAACCCCCCGTTCAGCCCGACCGCTGCGCCT
TATCCGGTAACTATCGTCTTGAGTCCAACCCGGTAAGACACGACTTATCGCCACTGGCAG
CAGCCACTGGTAACAGGATTAGCAGAGCGAGGTATGTAGGCGGTGCTACAGAGTTCTTGA
AGTGGTGGCCTAACTACGGCTACACTAGAAGGACAGTATTTGGTATCTGCGCTCTGCTGA
AGCCAGTTACCTTCGGAAAAAGAGTTGGTAGCTCTTGATCCGGCAAACAAACCACCGCTG
GTAGCGGTGGTTTTTTTGTTTGCAAGCAGCAGATTACGCGCAGAAAAAAAGGATCTCAAG
AAGATCCTTTGATCTTTTCTACGGGGTCTGACGCTCAGTGGAACGAAAACTCACGTTAAG
GGATTTTGGTCATGAGATTATCAAAAAGGATCTTCACCTAGATCCTTTTAAATTAAAAAT
GAAGTTTTAAATCAATCTAAAGTATATATGAGTAAACTTGGTCTGACAGTTACCAATGCT
TAATCAGTGAGGCACCTATCTCAGCGATCTGTCTATTTCGTTCATCCATAGTTGCCTGAC
TCCCCGTCGTGTAGATAACTACGATACGGGAGGGCTTACCATCTGGCCCCAGTGCTGCAA
TGATACCGCGAGACCCACGCTCACCGGCTCCAGATTTATCAGCAATAAACCAGCCAGCCG
GAAGGGCCGAGCGCAGAAGTGGTCCTGCAACTTTATCCGCCTCCATCCAGTCTATTAATT
GTTGCCGGGAAGCTAGAGTAAGTAGTTCGCCAGTTAATAGTTTGCGCAACGTTGTTGCCA
TTGCTACAGGCATCGTGGTGTCACGCTCGTCGTTTGGTATGGCTTCATTCAGCTCCGGTT
CCCAACGATCAAGGCGAGTTACATGATCCCCCATGTTGTGCAAAAAAGCGGTTAGCTCCT
TCGGTCCTCCGATCGTTGTCAGAAGTAAGTTGGCCGCAGTGTTATCACTCATGGTTATGG
CAGCACTGCATAATTCTCTTACTGTCATGCCATCCGTAAGATGCTTTTCTGTGACTGGTG
AGTACTCAACCAAGTCATTCTGAGAATAGTGTATGCGGCGACCGAGTTGCTCTTGCCCGG
CGTCAATACGGGATAATACCGCGCCACATAGCAGAACTTTAAAAGTGCTCATCATTGGAA
AACGTTCTTCGGGGCGAAAACTCTCAAGGATCTTACCGCTGTTGAGATCCAGTTCGATGT
AACCCACTCGTGCACCCAACTGATCTTCAGCATCTTTTACTTTCACCAGCGTTTCTGGGT
GAGCAAAAACAGGAAGGCAAAATGCCGCAAAAAAGGGAATAAGGGCGACACGGAAATGTT
GAATACTCATACTCTTCCTTTTTCAATATTATTGAAGCATTTATCAGGGTTATTGTCTCA
TGAGCGGATACATATTTGAATGTATTTAGAAAAATAAACAAATAGGGGTTCCGCGCACAT
TTCCCCGAAAAGTGCCACCTAAATTGTAAGCGTTAATATTTTGTTAAAATTCGCGTTAAA
TTTTTGTTAAATCAGCTCATTTTTTAACCAATAGGCCGAAATCGGCAAAATCCCTTATAA
ATCAAAAGAATAGACCGAGATAGGGTTGAGTGTTGTTCCAGTTTGGAACAAGAGTCCACT
ATTAAAGAACGTGGACTCCAACGTCAAAGGGCGAAAAACCGTCTATCAGGGCGATGGCCC
ACTACGTGAACCATCACCCTAATCAAGTTTTTTGGGGTCGAGGTGCCGTAAAGCACTAAA
TCGGAACCCTAAAGGGAGCCCCCGATTTAGAGCTTGACGGGGAAAGCCGGCGAACGTGGC
GAGAAAGGAAGGGAAGAAAGCGAAAGGAGCGGGCGCTAGGGCGCTGGCAAGTGTAGCGGT
CACGCTGCGCGTAACCACCACACCCGCCGCGCTTAATGCGCCGCTACAGGGCGCGTCCCA
TTCGCCATTCAGGCTGCGCAACTGTTGGGAAGGGCGATCGGTGCGGGCCTCTTCGCTATT
ACGCCAGCTGGCGAAAGGGGGATGTGCTGCAAGGCGATTAAGTTGGGTAACGCCAGGGTT
TTCCCAGTCACGACGTTGTAAAACGACGGCCAGTGAGCGCGCGTAATACGACTCACTATA
GGGCGAATTGGAGCTCCACCGCGGTGGCGGCCGCTCTAGAACTAGTGTATTGTCAAATCA
TCTTATATGTGCATCTATATAACGAATTTATAATGACTAAGATATATATATTATAAATTG
TTAATATTAAACTTTTTGGATATTTTTTTATGTTTTTAATATATATATTTTATTGGCTTA
AAATATATATTCGATATATCAAAGTATTATTATAAGATTAATACTAGGTTATTGATATAT
ATATATTTATAAGAACCTATATATGTTGTAACACATTAGTTTACATTGCTAATTAACCAA
TTGTTTGAACTATAATTAAATATATAAAAGAAAAAATGGGAAATCCTAATTTGTTAAAAA
TTTTTTTATTTATCTTTTTAATATGGATATGCTTATATTATAATAATGAGGTAAATAATT
AAATATATATTTTTTTAAATATTTTTTGTAATATTATTTTTTTATGTTATACGCACCTAT
TGAAGCCAATTTTGTTTACTTTCCACAAACATGATCACTCAACAATTGTAGTTGACGATA
TAAATAGAAATATGCTTATATATTAAACTATTTTGTGAACGATTTATGCACCCATATATA
TATACATATATTGTCATTTTTTTATTTTAGAATGGAATTAATAAATCGCTGAATAAAGTA
GAGAGTACAAATGGTTTATCAAATAGACAGTTTTATCGGATTTTAGAAGAAACAGAATGC
AAAAAAAAAAAACATTCAAAAAAAGTTAAGGAATCAAATAAGAAAAGTGATAATGGAAAA
AATATAATCAAATTAACAATTGACATTTCTCATGATAAATTAATTGCAACAACACTTATT
TTTGGATCATTGTTAATGTTATTAAATGTAGGATTCACTATGTTTATGTTTCTAAATAAA
TTTCGAAATGAACTTCCAGGATCCGGTGGAGGTGGAAGATCCATGGTGAGCAAGGGCGAG
GAGGATAACATGGCCATCATCAAGGAGTTCATGCGCTTCAAGGTGCACATGGAGGGCTCC
GTGAACGGCCACGAGTTCGAGATCGAGGGCGAGGGCGAGGGCCGCCCCTACGAGGGCACC
CAGACCGCCAAGCTGAAGGTGACCAAGGGTGGCCCCCTGCCCTTCGCCTGGGACATCCTG
TCCCCTCAGTTCATGTACGGCTCCAAGGCCTACGTGAAGCACCCCGCCGACATCCCCGAC
TACTTGAAGCTGTCCTTCCCCGAGGGCTTCAAGTGGGAGCGCGTGATGAACTTCGAGGAC
GGCGGCGTGGTGACCGTGACCCAGGACTCCTCCCTGCAGGACGGCGAGTTCATCTACAAG
GTGAAGCTGCGCGGCACCAACTTCCCCTCCGACGGCCCCGTAATGCAGAAGAAGACCATG
GGCTGGGAGGCCTCCTCCGAGCGGATGTACCCCGAGGACGGCGCCCTGAAGGGCGAGATC
AAGCAGAGGCTGAAGCTGAAGGACGGCGGCCACTACGACGCTGAGGTCAAGACCACCTAC
AAGGCCAAGAAGCCCGTGCAGCTGCCCGGCGCCTACAACGTCAACATCAAGTTGGACATC
ACCTCCCACAACGAGGACTACACCATCGTGGAACAGTACGAACGCGCCGAGGGCCGCCAC
TCCACCGGCGGCATGGACGAGCTGTACAAGTAATCTAGAATTATTAATATATATGAATAT
ATATACATCGTTGTATGCGTAATATACGTGTAACTGTTTTTTTTAATTTTTATATTAACT
TTTCCTTATACATTTTAATGCCATTAACTAATAATATTTTACTTGAAAAAATTCTAAATT
TTTATAACTCTAATATAAAATCTGTGAATAATAAAAAATTAGCATAAAGTACGAATTAAA
CATCGAATATTTGTTTTAAAGGGGCACACATTATGTGCATATAATTTTATTCTTTGAATT
TAATTTTTTGCTTATAATTAGATTAAATTAATATAATTTTTTTATTGAAAATAAAAACTT
ATTTTAAAAAATTAACAAATGGAAGTTAATGAAAAATACGAAAAAAGTAAAGGGTTAATT
CTTATATGGTCGGTACCGAGCTCGAATTCGATATCGCTAGCCTCGAGGGTACCCATCGAA
ATTGAAGGAAAAAACATCATTTGTGTTTATATGTTTTTTTTTAATTTTTCAACTTTTTTT
ATTTAACTATGCTAATCATATTATAAACATTTTAAAAATCAAGTCCAACTATTTATGAAT
CATTGAAGAGACAACATAATTATAACAGTAAAAATAATAATAATAATAATAATTTTGTCT
TATTGTTTCAAACATATTATGTTCCATAAATATCTAAATTTTTAATATGGTCATAAATTC
ATAGGATTTAATTTATATAAAATATGGCTATTCATACTAGCCATTTTATGTGTGTAAAGT
TTTTTTTTTCTGAACAAAAAATAAAATGTTCATAATTTTAGCTATTTACATGCATGTGCA
TGCACAAAAAAAAATATGCACACAACATACACATTTTTACAGTTATAAATACAATCAATT
GGTATAAATATATAAGTAAGAAAAACGGGATCCTTAATCATTCTTCTCATATACTTCAAA
TTTGTACTTAATGCCTTTCTCCTCCTGGACATCAGAGAGAACACCTGGGTATTCTGGCAG
AAGTTTATATTTCTCCAAATCAATTTCTGGAAAAAACGTGTCACTTTCAAAGTCTTGCAT
GATCCTTGTCACAAATAGTTTAAGATGGCCTGGGTGATTCATGGCTTCCTTATAAACAGA
ACTGCCACCAACTATCCAGACCATGTCTACTTTATTTGCTAATTCTGGTTGTTCAGTAAG
TTTTAAGGCATCATCTAGACTTCTGGAAAGAAAATGAGCTCCTTGTGGAGGTTCCTTGAG
TTCTCTGCTGAGAACTAAATTAATTCTACCCTTTAAAGGTCGATTCTTCTCAGGAATGGA
GAACCAGGTCTTCTTACCCATAATCACCAGATTCTGTTTACCTTCTACTGAAGAGGTTGT
GGTCATTCTCTGGAAATATCTAAATTCGTTCCTGAGCGGTGGCCAGGGCAGGTCCCCGTT
CTTGCCGATGCCCATGTTCTGGGACACAGCGACGATGCAGTTTAGCGAACCAACCATGGA
TCCCCCTATGTTTTATAAAATTTTTTATTTATTTATAAGCAAATATATATTTTTATATAT
TTATACACAAAGTATATATATTTTAGGAAATATTAAAATAAATTTAAATTTTGAAGGCAA
ATGAAAAAATTACGTTTTTTGTTTCACAACAATTATATTTATTTTTTGAATTAATTTACA
TATTATTTTATATTAAAATTATAAAATAAAAAAAACATTACATATATTAATTATATTATT
ATATGAGGATTTTTTTATAACATAGAAATATAAATAATTACGACTAGTTAATAAAGGGCA
CTTTAATTTTTATGCATAATTAGCTTATATTTAGTGCATATTTATTTTTATACATATTTT
TATATATTTTTTGATTTACATGGCGTTTTATGTTTATATGTATAACATTATATGCATAGT
ATTTTTTTGAAAATTTAATGGTATTTCAGTGGATGAAAATATTACTGGTGCTTTGAGGGG
TGAGCATTTAAAGCACAATATCTAGGATACTACAAAAAATACTTAAAGTATGAATATTAA
ATTGTAAACTTAAGCATAAAGAGCTCGAAAAGAATTAAGCTGGGCTGCAGAGGCCTGTTA
ACAGATCTATCGATCCGCGGGGGCCCAAGCT
Restriction sites to linearize plasmid BclI
Selectable marker used to select the mutant parasitetgdhfr
Promoter of the selectable markerpbdhfr
Selection (positive) procedurepyrimethamine
Selection (negative) procedureNo
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 1actagtGTATTGTCAAATCATCTTATATGTGC
Additional information primer 17124 (SpeI); PBANKA_083640 forward
Sequence Primer 2ggatccTGGAAGTTCATTTCGAAATTTATTTAG
Additional information primer 27125 (BamHI); PBANKA_083640 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