RMgmDB - Rodent Malaria genetically modified Parasites

Summary

RMgm-179
Malaria parasiteP. berghei
Genotype
DisruptedGene model (rodent): PBANKA_1117800; Gene model (P.falciparum): PF3D7_0618600; Gene product: rhomboid protease ROM10 (ROM10)
PhenotypeNo phenotype has been described
Last modified: 16 March 2013, 12:05
  *RMgm-179
Successful modificationThe parasite was generated by the genetic modification
The mutant contains the following genetic modification(s) Gene disruption
Reference (PubMed-PMID number) Reference 1 (PMID number) : 23490234
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. Lin, G.R. Mair, C.J. Janse, S.M. Khan
Name Group/DepartmentLeiden Malaria Research Group
Name InstituteLeiden University Medical Center (LUMC)
CityLeiden
CountryThe Netherlands
Name of the mutant parasite
RMgm numberRMgm-179
Principal name468cl2; 468cl3
Alternative name∆rom10
Standardized name
Is the mutant parasite cloned after genetic modificationYes
Phenotype
Asexual blood stageNot different from wild type
Gametocyte/GameteNot different from wild type
Fertilization and ookineteNot different from wild type
OocystNot different from wild type
SporozoiteNot different from wild type
Liver stageNot different from wild type
Additional remarks phenotype

Mutant/mutation
The mutant lacks expression of rhomboid protease ROM10.

Protein (function)
Rhomboid proteins are intra-membrane proteases that play a role in multiple processes. They belong to a family of serine proteases that cleave cell-surface proteins within their transmembrane domains. The Plasmodium  genome encodes a total of 8 rhomboid proteases (ROM1, 3, 4, 6, 7, 8, 9 and 10).

Phenotype
The phenotype analyses indicate that ROM10 is not essential throughout the complete life cycle.

Additional information
ROM10 has been detected in proteomes of P. falciparum asexual blood stages and gametocytes.

Other mutants
RMgm-176: A P. berghei mutant lacking expression of rhomboid protease ROM1 (partial disruption)
RMgm-177: A P. berghei mutant lacking expression of rhomboid protease ROM1 (partial disruption)
RMgm-761: A P. berghei mutant lacking expression of rhomboid protease ROM1 (complete disruption)
RMgm-178: A P. berghei mutant lacking expression of rhomboid protease ROM3
RMgm-762: A P. berghei mutant lacking expression of rhomboid protease ROM9

RMgm-187: Unsuccessful attempts to disrupt P. berghei rhomboid protease rom4
RMgm-758: Unsuccessful attempts to disrupt P. berghei rhomboid protease rom6
RMgm-759: Unsuccessful attempts to disrupt P. berghei rhomboid protease rom7
RMgm-760: Unsuccessful attempts to disrupt P. berghei rhomboid protease rom8

RMgm-763: A P. berghei mutant expressing  a (C-terminal) GFP-tagged version of rhomboid protease ROM3
RMgm-764: A P. berghei mutant expressing  a (C-terminal) mCherry-tagged version of rhomboid protease ROM4

RMgm-659: A P. y. yoelii 17XNL mutant lacking expression of rhomboid protease ROM1
RMgm-660: A P. y. yoelii 17XNL mutant expressing  a (N-terminal) HA-tagged version of rhomboid protease ROM1


  Disrupted: Mutant parasite with a disrupted gene
Details of the target gene
Gene Model of Rodent Parasite PBANKA_1117800
Gene Model P. falciparum ortholog PF3D7_0618600
Gene productrhomboid protease ROM10
Gene product: Alternative nameROM10
Details of the genetic modification
Inducable system usedNo
Additional remarks inducable system
Type of plasmid/construct usedPlasmid double 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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CTAGTAACGGCCGCCAGTGTGCTGGAATTCGGCAAGGGCCCTCACATTTTTCCCATCAGA
GCTTTCCGTTATTATTTCTCGCTTTTAATGTATTGTCCCGTTTTGATGTTTTCACATATT
TACTAACTTGTTTTACGAATTAAATACATTATTGCTAGTTTACAGTTTTTTATTTGTTTT
ATTGTTCCGTATCTTTTGTAAATGAATAATCTATTATATTAAAATTGAGTTTAGAATATT
TACGATTCTACATAAAATAATAAAAAAGCAAAAATGCGATTTATCATGAATTATCAAAAT
GTTCCAAATGACGAGGTATTTGATGACTCCGATTTGTTACTTTTAAAATATGCACGATCT
TTTTATAAAACTTTTAAGAAGAATATTCAATTTATTCAAATCTTATTCCTTTCTTATAAG
CTTCATTATGTTACCATAATCTTTTCCATATTCTTATATATATTTTTTTTGGGATATGAC
ATTTATTTTTTTAATAAATACAACCCTTTATTTATTAGAGAGTAAACCAATATTTACCAT
ATATATATATATAAATAAAAAAAAATTATAATTATTACTTGGGCATGTATAGACAGAATC
GATAATTAAGCCGAATTCTGCAGATATCCATCACACTGGCGGCCGCCACCGCGGTGGAGC
TCCAATTCGCCCTATAGTGAGTCGTATTACGCGCGCTCACTGGCCGTCGTTTTACAACGT
CGTGACTGGGAAAACCCTGGCGTTACCCAACTTAATCGCCTTGCAGCACATCCCCCTTTC
GCCAGCTGGCGTAATAGCGAAGAGGCCCGCACCGATCGCCCTTCCCAACAGTTGCGCAGC
CTGAATGGCGAATGGGACGCGCCCTGTAGCGGCGCATTAAGCGCGGCGGGTGTGGTGGTT
ACGCGCAGCGTGACCGCTACACTTGCCAGCGCCCTAGCGCCCGCTCCTTTCGCTTTCTTC
CCTTCCTTTCTCGCCACGTTCGCCGGCTTTCCCCGTCAAGCTCTAAATCGGGGGCTCCCT
TTAGGGTTCCGATTTAGTGCTTTACGGCACCTCGACCCCAAAAAACTTGATTAGGGTGAT
GGTTCACGTAGTGGGCCATCGCCCTGATAGACGGTTTTTCGCCCTTTGACGTTGGAGTCC
ACGTTCTTTAATAGTGGACTCTTGTTCCAAACTGGAACAACACTCAACCCTATCTCGGTC
TATTCTTTTGATTTATAAGGGATTTTGCCGATTTCGGCCTATTGGTTAAAAAATGAGCTG
ATTTAACAAAAATTTAACGCGAATTTTAACAAAATATTAACGCTTACAATTTAGGTGGCA
CTTTTCGGGGAAATGTGCGCGGAACCCCTATTTGTTTATTTTTCTAAATACATTCAAATA
TGTATCCGCTCATGAGACAATAACCCTGATAAATGCTTCAATAATATTGAAAAAGGAAGA
GTATGAGTATTCAACATTTCCGTGTCGCCCTTATTCCCTTTTTTGCGGCATTTTGCCTTC
CTGTTTTTGCTCACCCAGAAACGCTGGTGAAAGTAAAAGATGCTGAAGATCAGTTGGGTG
CACGAGTGGGTTACATCGAACTGGATCTCAACAGCGGTAAGATCCTTGAGAGTTTTCGCC
CCGAAGAACGTTTTCCAATGATGAGCACTTTTAAAGTTCTGCTATGTGGCGCGGTATTAT
CCCGTATTGACGCCGGGCAAGAGCAACTCGGTCGCCGCATACACTATTCTCAGAATGACT
TGGTTGAGTACTCACCAGTCACAGAAAAGCATCTTACGGATGGCATGACAGTAAGAGAAT
TATGCAGTGCTGCCATAACCATGAGTGATAACACTGCGGCCAACTTACTTCTGACAACGA
TCGGAGGACCGAAGGAGCTAACCGCTTTTTTGCACAACATGGGGGATCATGTAACTCGCC
TTGATCGTTGGGAACCGGAGCTGAATGAAGCCATACCAAACGACGAGCGTGACACCACGA
TGCCTGTAGCAATGGCAACAACGTTGCGCAAACTATTAACTGGCGAACTACTTACTCTAG
CTTCCCGGCAACAATTAATAGACTGGATGGAGGCGGATAAAGTTGCAGGACCACTTCTGC
GCTCGGCCCTTCCGGCTGGCTGGTTTATTGCTGATAAATCTGGAGCCGGTGAGCGTGGGT
CTCGCGGTATCATTGCAGCACTGGGGCCAGATGGTAAGCCCTCCCGTATCGTAGTTATCT
ACACGACGGGGAGTCAGGCAACTATGGATGAACGAAATAGACAGATCGCTGAGATAGGTG
CCTCACTGATTAAGCATTGGTAACTGTCAGACCAAGTTTACTCATATATACTTTAGATTG
ATTTAAAACTTCATTTTTAATTTAAAAGGATCTAGGTGAAGATCCTTTTTGATAATCTCA
TGACCAAAATCCCTTAACGTGAGTTTTCGTTCCACTGAGCGTCAGACCCCGTAGAAAAGA
TCAAAGGATCTTCTTGAGATCCTTTTTTTCTGCGCGTAATCTGCTGCTTGCAAACAAAAA
AACCACCGCTACCAGCGGTGGTTTGTTTGCCGGATCAAGAGCTACCAACTCTTTTTCCGA
AGGTAACTGGCTTCAGCAGAGCGCAGATACCAAATACTGTCCTTCTAGTGTAGCCGTAGT
TAGGCCACCACTTCAAGAACTCTGTAGCACCGCCTACATACCTCGCTCTGCTAATCCTGT
TACCAGTGGCTGCTGCCAGTGGCGATAAGTCGTGTCTTACCGGGTTGGACTCAAGACGAT
AGTTACCGGATAAGGCGCAGCGGTCGGGCTGAACGGGGGGTTCGTGCACACAGCCCAGCT
TGGAGCGAACGACCTACACCGAACTGAGATACCTACAGCGTGAGCTATGAGAAAGCGCCA
CGCTTCCCGAAGGGAGAAAGGCGGACAGGTATCCGGTAAGCGGCAGGGTCGGAACAGGAG
AGCGCACGAGGGAGCTTCCAGGGGGAAACGCCTGGTATCTTTATAGTCCTGTCGGGTTTC
GCCACCTCTGACTTGAGCGTCGATTTTTGTGATGCTCGTCAGGGGGGCGGAGCCTATGGA
AAAACGCCAGCAACGCGGCCTTTTTACGGTTCCTGGCCTTTTGCTGGCCTTTTGCTCACA
TGTTCTTTCCTGCGTTATCCCCTGATTCTGTGGATAACCGTATTACCGCCTTTGAGTGAG
CTGATACCGCTCGCCGCAGCCGAACGACCGAGCGCAGCGAGTCAGTGAGCGAGGAAGCGG
AAGAGCGCCCAATACGCAAACCGCCTCTCCCCGCGCGTTGGCCGATTCATTAATGCAGCT
GGCACGACAGGTTTCCCGACTGGAAAGCGGGCAGTGAGCGCAACGCAATTAATGTGAGTT
AGCTCACTCATTAGGCACCCCAGGCTTTACACTTTATGCTTCCGGCTCGTATGTTGTGTG
GAATTGTGAGCGGATAACAATTTCACACAGGAAACAGCTATGACCATGATTACGCCAAGC
TCGAAATTAACCCTCACTAAAGGGAACAAAAGCTGGTACCGGGCCCCCCCTCGAGGTCGA
CGGTATCGATAAGCTTGCATGCCTGCAGGTCAACAATAAATAATAAATAAATATTGTGGA
AATAAAATAACATATAATTATTTTTAATACATTGATTTCCCTTTTATTTTTTTAAATTTC
ATTGATATAAAAATATATAATAATAACATATATGATTTCAAATTAATCTTTTCAAAAATG
GTGTTTATTTTTGTATGTTTGTGTATGAATTAATCACATAACACATCTATTAAATTGAGT
TGGTAATATAGACACAAATAAATATATATATTTTTATAGCTTAAAAGTGTGTTATGAATA
TTTTAAGCATATTTTCTTTTTCTTTGGATTGTGTAAAATGAACTCATATAATGCGTTTTT
TTGTTTTTGTTATTTTGTCATTTTGTTATTTTGCTATTTTATGGATTAATTTTTGTTTAT
AAAATGGGAAATAATTTTAACATATTTAAATAAATGGAGAAAAAATATAAAATAATTATA
AAAAAAAGTTAATACACATTTTTTCCTGTTATAGACCTTATATTTATTTATCCATATATA
TATATATATATATATATATATATATATACATACCAAGTGAATTAAGAGGAAAGCTAATTT
ATTATTCAGAATAATATATGAACTATATATAATTTTTATTATTTTGGTGTATATTAATCT
GTCTATATGCATACATGCAATAATTTATCGACTTATATATCAAATAACATAAAATAGAAG
TGTTTTAAATTATGGATATATGCTCAATATTCATTTTTTTTAATAAGTTAGCTATATTTA
AATTATACATTTTATATATGGTCTCTTTTTTTTTTAAATATTATTTAAGTGATCATGAAA
ATATAAATAATTTTTTTTTATTTAATATCCTTTTGCTTGCATGTGGTAAATGGAAATTTG
GATGTGTTTTGAARGTTCGGATATAGTTGTATGGACATATAATATATTTTGTGAAAAATT
GGTTTTATGTTTATACTTATGCCAATACTTTTTGAGTAAAACAAAGCAAGTGCTTATAAA
TAATTAAAGCCAATTTTATAATATATATTTTTTTATTTAATTTGAATTTAGTAGTATAAT
TTTTTATGGTAAGTGCTCAAAGAGAGTTGCTTATAAAGTATGGTTTGTTTCTTTTTCGCC
ATTTTGAATTACACATTAAAAATATATAGATACATATATTATAATATGAAATCATTAATA
ATTTAGGGAAATTCTACAAATTTAAAAACGAATAAAATAATTGTTTTTCATCATGCCATA
ACACAATATTGATATATACATGTACAAACATTTTTTTTATTTGGAAAATATAAATTATAT
AAAAAAAAATGTATAGTATACAAAATGAGCATATTCACACGGGGTGGACGTTCATTTTTT
CATTTTTCCCCTGTTTTTTATGAGTATATGATAAAATTTTATGAACATTTACACAAAATG
AAAATGGATATATAGGAAAAATGGAGCGGTATTTCATTTATCTTTGATTGTCATTTGGAT
ATTATATTACCYTGGGTAGGCAATTAAAAATGTTAAATAACAATTTAAGGAAATTATATT
TTATATATTAAAATTAACACTGTATTATATGATTCGCTTATAAAAGCCACTCTTTCCCCA
TGCAAAGCTGTTTAATATCAATTTTAACAAATTACACACATGTTAATATATTTATATATA
TAATTTATATATTTATAATTTATATATTTATATTTTTATTATTTATATATTTATTATTTA
TTGTGTGTGTCAATTCGGGTAGGATATACCTCTTTTTTATTGTTTAAAGCGATTTGTATT
CTAAAATATAAAGRATTTGAAAAAGAGAAAGATAGAATATGATCCCATCATATATAGCCC
TATAATTTTTATTTAGCAGCGAATTAATTTTTCTATTAAGTTTATGTGTAATTAAAATAA
CGGAATATATATAATACAATAAAAAAGTGCATAAATTAAAATTTTTTCAATTAAATTTTT
TTTTTTAAGGGGTTATATAATATTAAATATATAAAATACGATTATATATTTTTGCTACAA
TTTTTTATATTAAGATATAAATAGTAAATAAATGGTATTATATGGCATGTAATATATAAA
TTTTTTCCAATTTTTATTTTATATACACTTTTCCTTTTTTTGTCATAAAACTTAAACAAT
TTACACATTCATTTTAAAAATTGACTATTTGTTTCAACATTTTTTGAGTTTCCGTTTTAT
AATAGTATTTTCATTTGTATATTGCTTATATATATAAATACACACCTAAATGTTACAAAG
GATCAATGCATAAACCGGTGTGTCTGGTCGTCGCGATGACCCCCAAGAGGGGCATCGGCA
TCAACAACGGCCTCCCGTGGCCCCACTTGACCACAGATTTCAAACACTTTTCTCGTGTGA
CAAAAACGACGCCCGAAGAAGCCAGTCGCCTGAACGGGTGGCTTCCCAGGAAATTTGCAA
AGACGGGCGACTCTGGACTTCCCTCTCCATCAGTCGGCAAGAGATTCAACGCCGTTGTCA
TGGGACGGAAAACCTGGGAAAGCATGCCTCGAAAGTTTAGACCCCTCGTGGACAGATTGA
ACATCGTCGTTTCCTCTTCCCTCAAAGAAGAAGACATTGCGGCGGAGAAGCCTCAAGCTG
AAGGCCAGCAGCGCGTCCGAGTCTGTGCTTCACTCCCAGCAGCTCTCAGCCTTCTGGAGG
AAGAGTACAAGGATTCTGTCGACCAGATTTTTGTCGTGGGAGGAGCGGGACTGTACGAGG
CAGCGCTGTCTCTGGGCGTTGCCTCTCACCTGTACATCACGCGTGTAGCCCGCGAGTTTC
CGTGCGACGTTTTCTTCCCTGCGTTCCCCGGAGATGACATTCTTTCAAACAAATCAACTG
CTGCGCAGGCTGCAGCTCCTGCCGAGTCTGTGTTCGTTCCCTTTTGTCCGGAGCTCGGAA
GAGAGAAGGACAATGAAGCGACGTATCGACCCATCTTCATTTCCAAGACCTTCTCAGACA
ACGGGGTTCCCTACGACTTTGTGGTTCTCGAGAAGAGAAGGAAGACTGACGACGCAGCCA
CTGCGGAACCGAGCAACGCAATGAGCTCCTTGACGTCCACGAGGGAGACAACTCCCGTGC
ACGGGTTGCAGGCTCCTTCTTCGGCCGCAGCCATTGCCCCGGTGTTGGCGTGGATGGACG
AAGAAGACCGGAAAAAACGCGAGCAAAAGGAACTGATTCGGGCCGTTCCGCATGTTCACT
TTAGAGGCCATGAAGAGTTCCAGTACCTTGATCTCATTGCCGACATTATTAACAATGGAA
GGACAATGGATGACCGAACGGGCGTTGGTGTCATCTCCAAATTCGGCTGCACTATGCGCT
ACTCGCTGGATCAGGCCTTTCCACTTCTCACCACAAAGCGTGTGTTCTGGAAAGGGGTCC
TCGAAGAGTTGCTGTGGTTCATTCGCGGCGACACGAACGCAAACCATCTTTCTGAGAAGG
GCGTGAAGATCTGGGACAAGAATGTGACACGCGAGTTCCTCGATTCGCGCAATCTCCCCC
ACCGAGAGGTCGGAGACATCGGCCCGGGCTACGGCTTCCAGTGGAGACACTTCGGCGCGG
CATACAAAGACATGCACACAGACTACACAGGGCAGGGCGTCGACCAGCTGAAGAATGTGA
TCCAGATGCTGAGAACGAATCCAACAGATCGTCGCATGCTCATGACTGCCTGGAATCCTG
CAGCGCTGGACGAAATGGCGCTGCCGCCTTGTCACTTGTTGTGCCAGTTCTACGTGAACG
ACCAGAAGGAGCTGTCGTGCATCATGTATCAGCGGTCGTGCGATGTCGGCCTCGGCGTCC
CCTTCAACATCGCTTCCTATTCGCTTTTGACGCTCATGGTTGCACACGTCTGCAACCTAA
AACCTAAGGAGTTCATTCACTTCATGGGGAACACGCATGTCTACACGAACCATGTCGAGG
CTTTAAAAGAGCAGCTGCGGAGAGAACCGAGACCGTTCCCCATTGTGAACATCCTCAACA
AGGAACGCATCAAGGAAATCGACGATTTCACCGCCGAGGATTTTGAGGTCGTGGGCTACG
TCCCGCACGGACGAATCCAGATGGAGATGGCTGTCTAGCGGAAATACAGAAGCTAGCTTT
GATCCCGTTTTTCTTACTTATATATTTATACCAATTGATTGTATTTATAACTGTAAAAAT
GTGTATGTTGTGTGCATATTTTTTTTTGTGCATGCACATGCATGTAAATAGCTAAAATTA
TGAACATTTTATTTTTTGTTCAGAAAAAAAAAACTTTACACACATAAAATGGCTAGTATG
AATAGCCATATTTTATATAAATTAAATCCTATGAATTTATGACCATATTAAAAATTTAGA
TATTTATGGAACATAATATGTTTGAAACAATAAGACAAAATTATTATTATTATTATTATT
TTTACTGTTATAATTATGTTGTCTCTTCAATGATTCATAAATAGTTGGACTTGATTTTTA
AAATGTTTATAATATGATTAGCATAGTTAAATAAAAAAAGTTGAAAAATTAAAAAAAAAC
ATATAAACACAAATGATGTTTTTTCCTTCAATTTCGATATCCATACGTAATATGTCTATG
CTATTTAAGAGGCGAATAAAACACCATTTTAGTTTCAGCATAAAATTAATGCATGCACAC
ATATGATAATGATAAATTAAAATATTAATAATTTATCATATTTATTAGTTGTGTACGATC
AATTTGACTATCCATTTTAAGGAAAAAAAAAACTTGTAAATTTAGTCAATTACATTCAAG
GCTTTTTGTGTATTTTCTCTAATATTGTTTATCGACTTTTGATAGCAAGCTTGTTCTTCT
TCTGTCAATATAAATTCGATTGGATGTGCCCCAGTTTTATTAATAATAGCTGTTGATCCA
GCATATAAATCTTTACAATTATATAGTCCACTTAAATAAACTGCACATGTAAATAATTGA
TTTTGATCGTGTAAATATGATTTTATCATTTTAATAATAGAAGCAGCCGGAGCAAATGCA
GCAGACGATTTAGCAAGCTTAATAATTTCACCACCCATATTCCTTGTCTTTTCTATAATA
TCACTTATATCTTTTTCTGACATATCATGATTTTTAATATAATCTAATAATGGTATACCC
GATATTGAACAATATCTTGGCAATGGCATCATAAGATCACCATGGCCTCCTAAAATAATA
GCATTAATATTTTCTGGGGAAACATTTAATTTTTCACTCAATAAATATCTAAATCTTGAT
GTATCTAATATACCTGCCATTCCACATATTTTTTCATATGGCAGATTACTATACTTGTGA
AAAACATTAACCATAACATCAAGAGGGTTTGTAACACATATAACAAAAGCATTTGGTGCA
TATTGTTTTACAGATTCAGCAACACTTTTTATTATTTTCCCATTTATACCAATTAAATCT
TCTCTGGACATTCCCTCCTTTCTTTGAACACCTGCAGTTATAACTGGATCCA
Restriction sites to linearize plasmid
Partial or complete disruption of the geneComplete
Additional remarks partial/complete disruption
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 1AAGGGCCCTCACATTTTTCCCATCAGAG
Additional information primer 1Pb 5' rho-8 sense (ApaI)
Sequence Primer 2TTATCGATTCTGTCTATACATGCCCAAG
Additional information primer 2Pb 5' rho-8 antisense (ClaI)
Sequence Primer 3AAGATATCCATACGTAATATGTCTATGC
Additional information primer 3Pb 3' rho-8 sense (EcoRV)
Sequence Primer 4TTGGATCCAGTTATAACTGCAGGTGTTC
Additional information primer 4Pb 3' rho-8 antisense (BamHI)
Sequence Primer 5
Additional information primer 5
Sequence Primer 6
Additional information primer 6