RMgmDB - Rodent Malaria genetically modified Parasites

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Summary

RMgm-252

Malaria parasite	P. berghei
Genotype
Disrupted	Gene model (rodent): PBANKA_1241700; Gene model (P.falciparum): PF3D7_0526900; Gene product: transmembrane emp24 domain-containing protein, putative
Phenotype	No phenotype has been described

Last modified: 6 July 2009, 10:37

*RMgm-252

Successful modification	The parasite was generated by the genetic modification
The mutant contains the following genetic modification(s)	Gene disruption
Reference (PubMed-PMID number)	Not published (yet)
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	S.M. Khan, C.J. Janse
Name Group/Department	Leiden Malaria Research Group
Name Institute	Leiden University Medical Center
City	Leiden
Country	The Netherlands
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Name of the mutant parasite
RMgm number	RMgm-252
Principal name	1108
Alternative name
Standardized name
Is the mutant parasite cloned after genetic modification	No
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Phenotype
Asexual blood stage	Not different from wild type
Gametocyte/Gamete	Not different from wild type
Fertilization and ookinete	Not different from wild type
Oocyst	Not tested
Sporozoite	Not tested
Liver stage	Not tested
Additional remarks phenotype	Mutant/mutation The mutant lacks expression of PB000809.01.0 Protein (function) Phenotype The phenotype of the (uncloned) mutant has not been analysed in detail. The mutant showed normal growth of the blood stages and normal production of gametocytes and ookinetes. The mutant can be obtained from the Leiden Malaria Research Group. Additional information Other mutants

Disrupted: Mutant parasite with a disrupted gene

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Details of the target gene

Gene Model of Rodent Parasite PBANKA_1241700

Gene Model P. falciparum ortholog PF3D7_0526900

Gene product transmembrane emp24 domain-containing protein, putative

Gene product: Alternative name

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Details of the genetic modification

Inducable system used No

Additional remarks inducable system

Type of plasmid/construct used Plasmid double cross-over

PlasmoGEM (Sanger) construct/vector used No

Modified PlasmoGEM construct/vector used No

Plasmid/construct map

Plasmid/construct sequence

GGGCCCCCCCTCGAGGTCGACGGTATCGATAAGCTTGCATGCCTGCAGGTCAACAATAAA

TAATAAATAAATATTGTGGAAATAAAATAACATATAATTATTTTTAATACATTGATTTCC

CTTTTATTTTTTTAAATTTCATTGATATAAAAATATATAATAATAACATATATGATTTCA

AATTAATCTTTTCAAAAATGGTGTTTATTTTTGTATGTTTGTGTATGAATTAATCACATA

ACACATCTATTAAATTGAGTTGGTAATATAGACACAAATAAATATATATATTTTTATAGC

TTAAAAGTGTGTTATGAATATTTTAAGCATATTTTCTTTTTCTTTGGATTGTGTAAAATG

AACTCATATAATGCGTTTTTTTGTTTTTGTTATTTTGTCATTTTGTTATTTTGCTATTTT

ATGGATTAATTTTTGTTTATAAAATGGGAAATAATTTTAACATATTTAAATAAATGGAGA

AAAAATATAAAATAATTATAAAAAAAAGTTAATACACATTTTTTCCTGTTATAGACCTTA

TATTTATTTATCCATATATATATATATATATATATATATATATATATACATACCAAGTGA

ATTAAGAGGAAAGCTAATTTATTATTCAGAATAATATATGAACTATATATAATTTTTATT

ATTTTGGTGTATATTAATCTGTCTATATGCATACATGCAATAATTTATCGACTTATATAT

CAAATAACATAAAATAGAAGTGTTTTAAATTATGGATATATGCTCAATATTCATTTTTTT

TAATAAGTTAGCTATATTTAAATTATACATTTTATATATGGTCTCTTTTTTTTTTAAATA

TTATTTAAGTGATCATGAAAATATAAATAATTTTTTTTTATTTAATATCCTTTTGCTTGC

ATGTGGTAAATGGAAATTTGGATGTGTTTTGAARGTTCGGATATAGTTGTATGGACATAT

AATATATTTTGTGAAAAATTGGTTTTATGTTTATACTTATGCCAATACTTTTTGAGTAAA

ACAAAGCAAGTGCTTATAAATAATTAAAGCCAATTTTATAATATATATTTTTTTATTTAA

TTTGAATTTAGTAGTATAATTTTTTATGGTAAGTGCTCAAAGAGAGTTGCTTATAAAGTA

TGGTTTGTTTCTTTTTCGCCATTTTGAATTACACATTAAAAATATATAGATACATATATT

ATAATATGAAATCATTAATAATTTAGGGAAATTCTACAAATTTAAAAACGAATAAAATAA

TTGTTTTTCATCATGCCATAACACAATATTGATATATACATGTACAAACATTTTTTTTAT

TTGGAAAATATAAATTATATAAAAAAAAATGTATAGTATACAAAATGAGCATATTCACAC

GGGGTGGACGTTCATTTTTTCATTTTTCCCCTGTTTTTTATGAGTATATGATAAAATTTT

ATGAACATTTACACAAAATGAAAATGGATATATAGGAAAAATGGAGCGGTATTTCATTTA

TCTTTGATTGTCATTTGGATATTATATTACCYTGGGTAGGCAATTAAAAATGTTAAATAA

CAATTTAAGGAAATTATATTTTATATATTAAAATTAACACTGTATTATATGATTCGCTTA

TAAAAGCCACTCTTTCCCCATGCAAAGCTGTTTAATATCAATTTTAACAAATTACACACA

TGTTAATATATTTATATATATAATTTATATATTTATAATTTATATATTTATATTTTTATT

ATTTATATATTTATTATTTATTGTGTGTGTCAATTCGGGTAGGATATACCTCTTTTTTAT

TGTTTAAAGCGATTTGTATTCTAAAATATAAAGRATTTGAAAAAGAGAAAGATAGAATAT

GATCCCATCATATATAGCCCTATAATTTTTATTTAGCAGCGAATTAATTTTTCTATTAAG

TTTATGTGTAATTAAAATAACGGAATATATATAATACAATAAAAAAGTGCATAAATTAAA

ATTTTTTCAATTAAATTTTTTTTTTTAAGGGGTTATATAATATTAAATATATAAAATACG

ATTATATATTTTTGCTACAATTTTTTATATTAAGATATAAATAGTAAATAAATGGTATTA

TATGGCATGTAATATATAAATTTTTTCCAATTTTTATTTTATATACACTTTTCCTTTTTT

TGTCATAAAACTTAAACAATTTACACATTCATTTTAAAAATTGACTATTTGTTTCAACAT

TTTTTGAGTTTCCGTTTTATAATAGTATTTTCATTTGTATATTGCTTATATATATAAATA

CACACCTAAATGTTACAAAGGATCAATGCATAAACCGGTGTGTCTGGTCGTCGCGATGAC

CCCCAAGAGGGGCATCGGCATCAACAACGGCCTCCCGTGGCCCCACTTGACCACAGATTT

CAAACACTTTTCTCGTGTGACAAAAACGACGCCCGAAGAAGCCAGTCGCCTGAACGGGTG

GCTTCCCAGGAAATTTGCAAAGACGGGCGACTCTGGACTTCCCTCTCCATCAGTCGGCAA

GAGATTCAACGCCGTTGTCATGGGACGGAAAACCTGGGAAAGCATGCCTCGAAAGTTTAG

ACCCCTCGTGGACAGATTGAACATCGTCGTTTCCTCTTCCCTCAAAGAAGAAGACATTGC

GGCGGAGAAGCCTCAAGCTGAAGGCCAGCAGCGCGTCCGAGTCTGTGCTTCACTCCCAGC

AGCTCTCAGCCTTCTGGAGGAAGAGTACAAGGATTCTGTCGACCAGATTTTTGTCGTGGG

AGGAGCGGGACTGTACGAGGCAGCGCTGTCTCTGGGCGTTGCCTCTCACCTGTACATCAC

GCGTGTAGCCCGCGAGTTTCCGTGCGACGTTTTCTTCCCTGCGTTCCCCGGAGATGACAT

TCTTTCAAACAAATCAACTGCTGCGCAGGCTGCAGCTCCTGCCGAGTCTGTGTTCGTTCC

CTTTTGTCCGGAGCTCGGAAGAGAGAAGGACAATGAAGCGACGTATCGACCCATCTTCAT

TTCCAAGACCTTCTCAGACAACGGGGTTCCCTACGACTTTGTGGTTCTCGAGAAGAGAAG

GAAGACTGACGACGCAGCCACTGCGGAACCGAGCAACGCAATGAGCTCCTTGACGTCCAC

GAGGGAGACAACTCCCGTGCACGGGTTGCAGGCTCCTTCTTCGGCCGCAGCCATTGCCCC

GGTGTTGGCGTGGATGGACGAAGAAGACCGGAAAAAACGCGAGCAAAAGGAACTGATTCG

GGCCGTTCCGCATGTTCACTTTAGAGGCCATGAAGAGTTCCAGTACCTTGATCTCATTGC

CGACATTATTAACAATGGAAGGACAATGGATGACCGAACGGGCGTTGGTGTCATCTCCAA

ATTCGGCTGCACTATGCGCTACTCGCTGGATCAGGCCTTTCCACTTCTCACCACAAAGCG

TGTGTTCTGGAAAGGGGTCCTCGAAGAGTTGCTGTGGTTCATTCGCGGCGACACGAACGC

AAACCATCTTTCTGAGAAGGGCGTGAAGATCTGGGACAAGAATGTGACACGCGAGTTCCT

CGATTCGCGCAATCTCCCCCACCGAGAGGTCGGAGACATCGGCCCGGGCTACGGCTTCCA

GTGGAGACACTTCGGCGCGGCATACAAAGACATGCACACAGACTACACAGGGCAGGGCGT

CGACCAGCTGAAGAATGTGATCCAGATGCTGAGAACGAATCCAACAGATCGTCGCATGCT

CATGACTGCCTGGAATCCTGCAGCGCTGGACGAAATGGCGCTGCCGCCTTGTCACTTGTT

GTGCCAGTTCTACGTGAACGACCAGAAGGAGCTGTCGTGCATCATGTATCAGCGGTCGTG

CGATGTCGGCCTCGGCGTCCCCTTCAACATCGCTTCCTATTCGCTTTTGACGCTCATGGT

TGCACACGTCTGCAACCTAAAACCTAAGGAGTTCATTCACTTCATGGGGAACACGCATGT

CTACACGAACCATGTCGAGGCTTTAAAAGAGCAGCTGCGGAGAGAACCGAGACCGTTCCC

CATTGTGAACATCCTCAACAAGGAACGCATCAAGGAAATCGACGATTTCACCGCCGAGGA

TTTTGAGGTCGTGGGCTACGTCCCGCACGGACGAATCCAGATGGAGATGGCTGTCTAGCG

GAAATACAGAAGCTAGCTTTGATCCCGTTTTTCTTACTTATATATTTATACCAATTGATT

GTATTTATAACTGTAAAAATGTGTATGTTGTGTGCATATTTTTTTTTGTGCATGCACATG

CATGTAAATAGCTAAAATTATGAACATTTTATTTTTTGTTCAGAAAAAAAAAACTTTACA

CACATAAAATGGCTAGTATGAATAGCCATATTTTATATAAATTAAATCCTATGAATTTAT

GACCATATTAAAAATTTAGATATTTATGGAACATAATATGTTTGAAACAATAAGACAAAA

TTATTATTATTATTATTATTTTTACTGTTATAATTATGTTGTCTCTTCAATGATTCATAA

ATAGTTGGACTTGATTTTTAAAATGTTTATAATATGATTAGCATAGTTAAATAAAAAAAG

TTGAAAAATTAAAAAAAAACATATAAACACAAATGATGTTTTTTCCTTCAATTTCGATAT

CGAATTCGCACATACATATGCGTATATATGCGCGTATATTATTATTATTGATAATATGCA

TTTTGTATATGAATTTTTTTTTTGAAAAATTTAAAGTGAAACACTTGTTTGAGTAAAAAA

TAAATTAATCTATGTATTTTAAAAAATAAAAAAAGATCTCCACATTTTACAATTAAAAAA

AAGTAAACGCGTATACATGCGTGTGATATTTGATGAGTAATTTAGACCAATTCAGACCAA

TTTAGACCAATTTAAAAAAAATTAAATAATAAAAATAGAAAAATAATATAAATAATAAAA

ATAGTAAAAATACCGAATGAAAAATAAATATACATTTGTAGCAGTTTAAAGAAAAAATGC

AAAAATAAAACGAAAAAAATGGAAAATAATTCGATAAATCAGCATTTATAAACTATTTTG

TCCTTTTTGACATAATAAATTGGTGCACCGTCCATCTCGTTCAAGGAATTTATAATTTTG

CGAAGGGTCGAGATATCCAACTTGGCTATCTGAGGCATTTTATCTTTGTTGCCCAAGATG

ATCTGAAGGAGGTGAAAAAATGAGAAAAATTAATGGGAAAAAAAATAGGCAAAGTGAAAG

TGAAAAAATAGGCATCTAGAGCGGCCGCCACCGCGGTGGAGCTCCAATTCGCCCTATAGT

GAGTCGTATTACAATTCACTGGCCGTCGTTTTACAACGTCGTGACTGGGAAAACCCTGGC

GTTACCCAACTTAATCGCCTTGCAGCACATCCCCCTTTCGCCAGCTGGCGTAATAGCGAA

GAGGCCCGCACCGATCGCCCTTCCCAACAGTTGCGCAGCCTGAATGGCGAATGGGACGCG

CCCTGTAGCGGCGCATTAAGCGCGGCGGGTGTGGTGGTTACGCGCAGCGTGACCGCTACA

CTTGCCAGCGCCCTAGCGCCCGCTCCTTTCGCTTTCTTCCCTTCCTTTCTCGCCACGTTC

GCCGGCTTTCCCCGTCAAGCTCTAAATCGGGGGCTCCCTTTAGGGTTCCGATTTAGTGCT

TTACGGCACCTCGACCCCAAAAAACTTGATTAGGGTGATGGTTCACGTAGTGGGCCATCG

CCCTGATAGACGGTTTTTCGCCCTTTGACGTTGGAGTCCACGTTCTTTAATAGTGGACTC

TTGTTCCAAACTGGAACAACACTCAACCCTATCTCGGTCTATTCTTTTGATTTATAAGGG

ATTTTGCCGATTTCGGCCTATTGGTTAAAAAATGAGCTGATTTAACAAAAATTTAACGCG

AATTTTAACAAAATATTAACGCTTACAATTTAGGTGGCACTTTTCGGGGAAATGTGCGCG

GAACCCCTATTTGTTTATTTTTCTAAATACATTCAAATATGTATCCGCTCATGAGACAAT

AACCCTGATAAATGCTTCAATAATATTGAAAAAGGAAGAGTATGAGTATTCAACATTTCC

GTGTCGCCCTTATTCCCTTTTTTGCGGCATTTTGCCTTCCTGTTTTTGCTCACCCAGAAA

CGCTGGTGAAAGTAAAAGATGCTGAAGATCAGTTGGGTGCACGAGTGGGTTACATCGAAC

TGGATCTCAACAGCGGTAAGATCCTTGAGAGTTTTCGCCCCGAAGAACGTTTTCCAATGA

TGAGCACTTTTAAAGTTCTGCTATGTGGCGCGGTATTATCCCGTATTGACGCCGGGCAAG

AGCAACTCGGTCGCCGCATACACTATTCTCAGAATGACTTGGTTGAGTACTCACCAGTCA

CAGAAAAGCATCTTACGGATGGCATGACAGTAAGAGAATTATGCAGTGCTGCCATAACCA

TGAGTGATAACACTGCGGCCAACTTACTTCTGACAACGATCGGAGGACCGAAGGAGCTAA

CCGCTTTTTTGCACAACATGGGGGATCATGTAACTCGCCTTGATCGTTGGGAACCGGAGC

TGAATGAAGCCATACCAAACGACGAGCGTGACACCACGATGCCTGTAGCAATGGCAACAA

CGTTGCGCAAACTATTAACTGGCGAACTACTTACTCTAGCTTCCCGGCAACAATTAATAG

ACTGGATGGAGGCGGATAAAGTTGCAGGACCACTTCTGCGCTCGGCCCTTCCGGCTGGCT

GGTTTATTGCTGATAAATCTGGAGCCGGTGAGCGTGGGTCTCGCGGTATCATTGCAGCAC

TGGGGCCAGATGGTAAGCCCTCCCGTATCGTAGTTATCTACACGACGGGGAGTCAGGCAA

CTATGGATGAACGAAATAGACAGATCGCTGAGATAGGTGCCTCACTGATTAAGCATTGGT

AACTGTCAGACCAAGTTTACTCATATATACTTTAGATTGATTTAAAACTTCATTTTTAAT

TTAAAAGGATCTAGGTGAAGATCCTTTTTGATAATCTCATGACCAAAATCCCTTAACGTG

AGTTTTCGTTCCACTGAGCGTCAGACCCCGTAGAAAAGATCAAAGGATCTTCTTGAGATC

CTTTTTTTCTGCGCGTAATCTGCTGCTTGCAAACAAAAAAACCACCGCTACCAGCGGTGG

TTTGTTTGCCGGATCAAGAGCTACCAACTCTTTTTCCGAAGGTAACTGGCTTCAGCAGAG

CGCAGATACCAAATACTGTCCTTCTAGTGTAGCCGTAGTTAGGCCACCACTTCAAGAACT

CTGTAGCACCGCCTACATACCTCGCTCTGCTAATCCTGTTACCAGTGGCTGCTGCCAGTG

GCGATAAGTCGTGTCTTACCGGGTTGGACTCAAGACGATAGTTACCGGATAAGGCGCAGC

GGTCGGGCTGAACGGGGGGTTCGTGCACACAGCCCAGCTTGGAGCGAACGACCTACACCG

AACTGAGATACCTACAGCGTGAGCTATGAGAAAGCGCCACGCTTCCCGAAGGGAGAAAGG

CGGACAGGTATCCGGTAAGCGGCAGGGTCGGAACAGGAGAGCGCACGAGGGAGCTTCCAG

GGGGAAACGCCTGGTATCTTTATAGTCCTGTCGGGTTTCGCCACCTCTGACTTGAGCGTC

GATTTTTGTGATGCTCGTCAGGGGGGCGGAGCCTATGGAAAAACGCCAGCAACGCGGCCT

TTTTACGGTTCCTGGCCTTTTGCTGGCCTTTTGCTCACATGTTCTTTCCTGCGTTATCCC

CTGATTCTGTGGATAACCGTATTACCGCCTTTGAGTGAGCTGATACCGCTCGCCGCAGCC

GAACGACCGAGCGCAGCGAGTCAGTGAGCGAGGAAGCGGAAGAGCGCCCAATACGCAAAC

CGCCTCTCCCCGCGCGTTGGCCGATTCATTAATGCAGCTGGCACGACAGGTTTCCCGACT

GGAAAGCGGGCAGTGAGCGCAACGCAATTAATGTGAGTTAGCTCACTCATTAGGCACCCC

AGGCTTTACACTTTATGCTTCCGGCTCGTATGTTGTGTGGAATTGTGAGCGGATAACAAT

TTCACACAGGAAACAGCTATGACCATGATTACGCCAAGCTCGAAATTAACCCTCACTAAA

GGGAACAAAAGCTGGTACCCCGCGGGATAAACTTTTCATTGAATTTGGTATGCAAACTTG

ACCAAAGTTTATGCATTTTTTCTCTTTTATTTTTAGGAGAATAAAAAAAAAATTAATAGT

AAAGATATTTATTACAATATACTACGCATAAGACCAAGTGACCTATGGATTTAAGACTTA

GAAGAAGTAGAGTTTGAATATAAAATAAGAGTGTGCGAAAAAAAATTTATTAATTTTGTT

CAAGCTTAAAATTTTCCTAAAATAAATATAATACAATTAATAATAACTTAATAATTTTTT

TATTATGAAAGTTGTGATTATTAAAATTTTCAGTAGTAAATAAAATAATATATTTTAAAT

AAAGATTAAAATGCTATATTTAATTAATATAATTTTTTAAATTATGAAAGGAATAATATT

GATTGGATATTTATTATTTATAGCATGTTTAAGGTACC

Restriction sites to linearize plasmid

Partial or complete disruption of the gene Complete

Additional remarks partial/complete disruption

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	GGCCGGTACCCCGCGGGATAAACTTTTCATTGAATTTGGTATGC
Additional information primer 1	L3386 (Asp718/SacII); 5'
Sequence Primer 2	GGCCGGTACCTTAAACATGCTATAAATAATAAATATCC
Additional information primer 2	L3376 (Asp718); 5'
Sequence Primer 3	GGCCGAATTCGCACATACATATGCGTATATATGCGCG
Additional information primer 3	L3377 (EcoRI); 3'
Sequence Primer 4	GGCCTCTAGATGCCTATTTTTTCACTTTCACTTTGCC
Additional information primer 4	L3378 (XbaI); 3'
Sequence Primer 5
Additional information primer 5
Sequence Primer 6
Additional information primer 6

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