RMgmDB - Rodent Malaria genetically modified Parasites

Summary

RMgm-108

Malaria parasite	P. berghei
Genotype
Disrupted	Gene model (rodent): PBANKA_1309600; Gene model (P.falciparum): PF3D7_1445800; Gene product: conserved Plasmodium membrane protein, unknown function
Phenotype	Oocyst; Sporozoite;

Last modified: 18 February 2009, 11:56

*RMgm-108

Successful modification	The 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) : 18974882
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 507cl1 (RMgm-7)
Other information parent line	P.berghei ANKA 507cl1 (RMgm-7) is a reference ANKA mutant line which expresses GFP under control of a constitutive promoter (PubMed: PMID: 16242190).
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The mutant parasite was generated by
Name PI/Researcher	E. Lasonder, C.J. Janse, H.G. Stunnenberg
Name Group/Department	Department of Molecular Biology
Name Institute	NCMLS, Radboud University Nijmegen
City	Nijmegen
Country	The Netherlands
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Name of the mutant parasite
RMgm number	RMgm-108
Principal name	802cl1; 838
Alternative name
Standardized name
Is the mutant parasite cloned after genetic modification	Yes
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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	Oocyst numbers in A. stephensi were comparable to wild type (ranging from 150–250 oocysts per mosquito). Sporozoite formation blocked inside the oocysts. No sporozoite formation was detectable within the oocysts by either fluorescence or phase-contrast microscopy.
Sporozoite	Sporozoite formation blocked inside the oocysts. No sporozoite formation was detectable within the oocysts by either fluorescence or phase-contrast microscopy. No sporozoites in salivary glands. No blood stage infection in C57BL/6 mice after feeding of infected mosquitoes on these mice.
Liver stage	Not tested
Additional remarks phenotype	Mutant/mutation The mutant lacks expression of PB101363.00.0 Protein (function) Unknown. The protein was detected in a proteome analysis of P. falciparum oocysts and sporozoites. It was highly abundant in oocyst-derived sporozoites and not detected in salivary gland sporozoites. It contains a signal peptide and a transmembrane region. Phenotype The phenotype analyses indicate a role of the protein in the formation of sporozoites within the oocysts. The mutant produces normal numbers of oocysts; however, no sporozoite formation was detectable within these oocysts.This early function in sporozoite development of this protein is in agreement with its presence in oocyst derived sporozoites and absence in salivary gland sporozoites.

Disrupted: Mutant parasite with a disrupted gene

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Details of the target gene

Gene Model of Rodent Parasite PBANKA_1309600

Gene Model P. falciparum ortholog PF3D7_1445800

Gene product conserved Plasmodium membrane protein, unknown function

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

GCGGATAACAATTTCACACAGGAAACAGCTATGACCATGATTACGCCAAGCTCGAAATTA

ACCCTCACTAAAGGGAACAAAAGCTGGTACCAAATAGTGCCTTTTTCGGTTTTTATGTTC

TCAAATACAATTTCATAGGTTTAATGTTGGTAGACAGGTAATTTTATAGTCAATATAATT

TAAAGCTAATATAATTTGTATAATATTTAACATAGGAACTTTTAAAAATATAATTATATT

TTTTTATACAATTTTTTAAATATATAAATCGTTGAAAAAGTATTGGAAAATAAAGATTGT

ATATATATACGCATGCACCCATGTAACTATAATGCAAGCACAAACGTTTATTCAATCTAA

AATATACATATATATATTGATATAAAAAAAAATAAAAATAAGCCCTATATTTCAAATTAC

TACTAAAATAAAGATATATCTTGGAAGCATAAATATATTATTTCAAGAAATATTTTTAGA

AGTCTTATAACCATTCTCAAGCTTGCATGCCTGCAGGTCAACAATAAATAATAAATAAAT

ATTGTGGAAATAAAATAACATATAATTATTTTTAATACATTGATTTCCCTTTTATTTTTT

TAAATTTCATTGATATAAAAATATATAATAATAACATATATGATTTCAAATTAATCTTTT

CAAAAATGGTGTTTATTTTTGTATGTTTGTGTATGAATTAATCACATAACACATCTATTA

AATTGAGTTGGTAATATAGACACAAATAAATATATATATTTTTATAGCTTAAAAGTGTGT

TATGAATATTTTAAGCATATTTTCTTTTTCTTTGGATTGTGTAAAATGAACTCATATAAT

GCGTTTTTTTGTTTTTGTTATTTTGTCATTTTGTTATTTTGCTATTTTATGGATTAATTT

TTGTTTATAAAATGGGAAATAATTTTAACATATTTAAATAAATGGAGAAAAAATATAAAA

TAATTATAAAAAAAAGTTAATACACATTTTTTCCTGTTATAGACCTTATATTTATTTATC

CATATATATATATATATATATATATATATATATATACATACCAAGTGAATTAAGAGGAAA

GCTAATTTATTATTCAGAATAATATATGAACTATATATAATTTTTATTATTTTGGTGTAT

ATTAATCTGTCTATATGCATACATGCAATAATTTATCGACTTATATATCAAATAACATAA

AATAGAAGTGTTTTAAATTATGGATATATGCTCAATATTCATTTTTTTTAATAAGTTAGC

TATATTTAAATTATACATTTTATATATGGTCTCTTTTTTTTTTAAATATTATTTAAGTGA

TCATGAAAATATAAATAATTTTTTTTTATTTAATATCCTTTTGCTTGCATGTGGTAAATG

GAAATTTGGATGTGTTTTGAARGTTCGGATATAGTTGTATGGACATATAATATATTTTGT

GAAAAATTGGTTTTATGTTTATACTTATGCCAATACTTTTTGAGTAAAACAAAGCAAGTG

CTTATAAATAATTAAAGCCAATTTTATAATATATATTTTTTTATTTAATTTGAATTTAGT

AGTATAATTTTTTATGGTAAGTGCTCAAAGAGAGTTGCTTATAAAGTATGGTTTGTTTCT

TTTTCGCCATTTTGAATTACACATTAAAAATATATAGATACATATATTATAATATGAAAT

CATTAATAATTTAGGGAAATTCTACAAATTTAAAAACGAATAAAATAATTGTTTTTCATC

ATGCCATAACACAATATTGATATATACATGTACAAACATTTTTTTTATTTGGAAAATATA

AATTATATAAAAAAAAATGTATAGTATACAAAATGAGCATATTCACACGGGGTGGACGTT

CATTTTTTCATTTTTCCCCTGTTTTTTATGAGTATATGATAAAATTTTATGAACATTTAC

ACAAAATGAAAATGGATATATAGGAAAAATGGAGCGGTATTTCATTTATCTTTGATTGTC

ATTTGGATATTATATTACCYTGGGTAGGCAATTAAAAATGTTAAATAACAATTTAAGGAA

ATTATATTTTATATATTAAAATTAACACTGTATTATATGATTCGCTTATAAAAGCCACTC

TTTCCCCATGCAAAGCTGTTTAATATCAATTTTAACAAATTACACACATGTTAATATATT

TATATATATAATTTATATATTTATAATTTATATATTTATATTTTTATTATTTATATATTT

ATTATTTATTGTGTGTGTCAATTCGGGTAGGATATACCTCTTTTTTATTGTTTAAAGCGA

TTTGTATTCTAAAATATAAAGRATTTGAAAAAGAGAAAGATAGAATATGATCCCATCATA

TATAGCCCTATAATTTTTATTTAGCAGCGAATTAATTTTTCTATTAAGTTTATGTGTAAT

TAAAATAACGGAATATATATAATACAATAAAAAAGTGCATAAATTAAAATTTTTTCAATT

AAATTTTTTTTTTTAAGGGGTTATATAATATTAAATATATAAAATACGATTATATATTTT

TGCTACAATTTTTTATATTAAGATATAAATAGTAAATAAATGGTATTATATGGCATGTAA

TATATAAATTTTTTCCAATTTTTATTTTATATACACTTTTCCTTTTTTTGTCATAAAACT

TAAACAATTTACACATTCATTTTAAAAATTGACTATTTGTTTCAACATTTTTTGAGTTTC

CGTTTTATAATAGTATTTTCATTTGTATATTGCTTATATATATAAATACACACCTAAATG

TTACAAAGGATCAATGCATAAACCGGTGTGTCTGGTCGTCGCGATGACCCCCAAGAGGGG

CATCGGCATCAACAACGGCCTCCCGTGGCCCCACTTGACCACAGATTTCAAACACTTTTC

TCGTGTGACAAAAACGACGCCCGAAGAAGCCAGTCGCCTGAACGGGTGGCTTCCCAGGAA

ATTTGCAAAGACGGGCGACTCTGGACTTCCCTCTCCATCAGTCGGCAAGAGATTCAACGC

CGTTGTCATGGGACGGAAAACCTGGGAAAGCATGCCTCGAAAGTTTAGACCCCTCGTGGA

CAGATTGAACATCGTCGTTTCCTCTTCCCTCAAAGAAGAAGACATTGCGGCGGAGAAGCC

TCAAGCTGAAGGCCAGCAGCGCGTCCGAGTCTGTGCTTCACTCCCAGCAGCTCTCAGCCT

TCTGGAGGAAGAGTACAAGGATTCTGTCGACCAGATTTTTGTCGTGGGAGGAGCGGGACT

GTACGAGGCAGCGCTGTCTCTGGGCGTTGCCTCTCACCTGTACATCACGCGTGTAGCCCG

CGAGTTTCCGTGCGACGTTTTCTTCCCTGCGTTCCCCGGAGATGACATTCTTTCAAACAA

ATCAACTGCTGCGCAGGCTGCAGCTCCTGCCGAGTCTGTGTTCGTTCCCTTTTGTCCGGA

GCTCGGAAGAGAGAAGGACAATGAAGCGACGTATCGACCCATCTTCATTTCCAAGACCTT

CTCAGACAACGGGGTTCCCTACGACTTTGTGGTTCTCGAGAAGAGAAGGAAGACTGACGA

CGCAGCCACTGCGGAACCGAGCAACGCAATGAGCTCCTTGACGTCCACGAGGGAGACAAC

TCCCGTGCACGGGTTGCAGGCTCCTTCTTCGGCCGCAGCCATTGCCCCGGTGTTGGCGTG

GATGGACGAAGAAGACCGGAAAAAACGCGAGCAAAAGGAACTGATTCGGGCCGTTCCGCA

TGTTCACTTTAGAGGCCATGAAGAGTTCCAGTACCTTGATCTCATTGCCGACATTATTAA

CAATGGAAGGACAATGGATGACCGAACGGGCGTTGGTGTCATCTCCAAATTCGGCTGCAC

TATGCGCTACTCGCTGGATCAGGCCTTTCCACTTCTCACCACAAAGCGTGTGTTCTGGAA

AGGGGTCCTCGAAGAGTTGCTGTGGTTCATTCGCGGCGACACGAACGCAAACCATCTTTC

TGAGAAGGGCGTGAAGATCTGGGACAAGAATGTGACACGCGAGTTCCTCGATTCGCGCAA

TCTCCCCCACCGAGAGGTCGGAGACATCGGCCCGGGCTACGGCTTCCAGTGGAGACACTT

CGGCGCGGCATACAAAGACATGCACACAGACTACACAGGGCAGGGCGTCGACCAGCTGAA

GAATGTGATCCAGATGCTGAGAACGAATCCAACAGATCGTCGCATGCTCATGACTGCCTG

GAATCCTGCAGCGCTGGACGAAATGGCGCTGCCGCCTTGTCACTTGTTGTGCCAGTTCTA

CGTGAACGACCAGAAGGAGCTGTCGTGCATCATGTATCAGCGGTCGTGCGATGTCGGCCT

CGGCGTCCCCTTCAACATCGCTTCCTATTCGCTTTTGACGCTCATGGTTGCACACGTCTG

CAACCTAAAACCTAAGGAGTTCATTCACTTCATGGGGAACACGCATGTCTACACGAACCA

TGTCGAGGCTTTAAAAGAGCAGCTGCGGAGAGAACCGAGACCGTTCCCCATTGTGAACAT

CCTCAACAAGGAACGCATCAAGGAAATCGACGATTTCACCGCCGAGGATTTTGAGGTCGT

GGGCTACGTCCCGCACGGACGAATCCAGATGGAGATGGCTGTCTAGCGGAAATACAGAAG

CTAGCTTTGATCCCGTTTTTCTTACTTATATATTTATACCAATTGATTGTATTTATAACT

GTAAAAATGTGTATGTTGTGTGCATATTTTTTTTTGTGCATGCACATGCATGTAAATAGC

TAAAATTATGAACATTTTATTTTTTGTTCAGAAAAAAAAAACTTTACACACATAAAATGG

CTAGTATGAATAGCCATATTTTATATAAATTAAATCCTATGAATTTATGACCATATTAAA

AATTTAGATATTTATGGAACATAATATGTTTGAAACAATAAGACAAAATTATTATTATTA

TTATTATTTTTACTGTTATAATTATGTTGTCTCTTCAATGATTCATAAATAGTTGGACTT

GATTTTTAAAATGTTTATAATATGATTAGCATAGTTAAATAAAAAAAGTTGAAAAATTAA

AAAAAAACATATAAACACAAATGATGTTTTTTCCTTCAATTTCGATATCGAATTCAGAAA

ATGCATTTTGAAGATTTAAAAAATATTTTTTGTCAAAATAAAGAAGATTTATGTACAAAT

TTTTATAAACTTATTTATTTTTGGATGAACATGAAAGACGATGAAATATATGATAGCCAA

TTTGTTCGAATTAAGACAGATTTTCAGAATAATATGAATAAATTATTACCTCAATTTTAT

ACAATAACTCCACATTTGTTCTATGAAAATTTTATAGTGGTATAGAAAATAACAAAATAT

AATAGTAATGTGTGTCTATAAGTGTTTAACTTTTCGGTTTGATATTATCATTTATTTTAT

TTTCATGTATATGCAGATGGACCCGTCTCATAACATAACGAGCAGTAGGATCGGATTTTT

TCTTCATAATTATCCAACAAATCAAGATAAAAACATACAATTATTAAAAAAAATAGAAAA

AATTGTAAAAAATAGCAATATCAAAGATGTATATTTTTATTCAGATACATATGTATTGTA

TAATCGCGGCCGCCACCGCGGTGGAGCTCCAATTCGCCCTATAGTGAGTCGTATTACGCG

CGCTCACTGGCCGTCGTTTTACAACGTCGTGACTGGGAAAACCCTGGCGTTACCCAACTT

AATCGCCTTGCAGCACATCCCCCTTTCGCCAGCTGGCGTAATAGCGAAGAGGCCCGCACC

GATCGCCCTTCCCAACAGTTGCGCAGCCTGAATGGCGAATGGGACGCGCCCTGTAGCGGC

GCATTAAGCGCGGCGGGTGTGGTGGTTACGCGCAGCGTGACCGCTACACTTGCCAGCGCC

CTAGCGCCCGCTCCTTTCGCTTTCTTCCCTTCCTTTCTCGCCACGTTCGCCGGCTTTCCC

CGTCAAGCTCTAAATCGGGGGCTCCCTTTAGGGTTCCGATTTAGTGCTTTACGGCACCTC

GACCCCAAAAAACTTGATTAGGGTGATGGTTCACGTAGTGGGCCATCGCCCTGATAGACG

GTTTTTCGCCCTTTGACGTTGGAGTCCACGTTCTTTAATAGTGGACTCTTGTTCCAAACT

GGAACAACACTCAACCCTATCTCGGTCTATTCTTTTGATTTATAAGGGATTTTGCCGATT

TCGGCCTATTGGTTAAAAAATGAGCTGATTTAACAAAAATTTAACGCGAATTTTAACAAA

ATATTAACGCTTACAATTTAGGTGGCACTTTTCGGGGAAATGTGCGCGGAACCCCTATTT

GTTTATTTTTCTAAATACATTCAAATATGTATCCGCTCATGAGACAATAACCCTGATAAA

TGCTTCAATAATATTGAAAAAGGAAGAGTATGAGTATTCAACATTTCCGTGTCGCCCTTA

TTCCCTTTTTTGCGGCATTTTGCCTTCCTGTTTTTGCTCACCCAGAAACGCTGGTGAAAG

TAAAAGATGCTGAAGATCAGTTGGGTGCACGAGTGGGTTACATCGAACTGGATCTCAACA

GCGGTAAGATCCTTGAGAGTTTTCGCCCCGAAGAACGTTTTCCAATGATGAGCACTTTTA

AAGTTCTGCTATGTGGCGCGGTATTATCCCGTATTGACGCCGGGCAAGAGCAACTCGGTC

GCCGCATACACTATTCTCAGAATGACTTGGTTGAGTACTCACCAGTCACAGAAAAGCATC

TTACGGATGGCATGACAGTAAGAGAATTATGCAGTGCTGCCATAACCATGAGTGATAACA

CTGCGGCCAACTTACTTCTGACAACGATCGGAGGACCGAAGGAGCTAACCGCTTTTTTGC

ACAACATGGGGGATCATGTAACTCGCCTTGATCGTTGGGAACCGGAGCTGAATGAAGCCA

TACCAAACGACGAGCGTGACACCACGATGCCTGTAGCAATGGCAACAACGTTGCGCAAAC

TATTAACTGGCGAACTACTTACTCTAGCTTCCCGGCAACAATTAATAGACTGGATGGAGG

CGGATAAAGTTGCAGGACCACTTCTGCGCTCGGCCCTTCCGGCTGGCTGGTTTATTGCTG

ATAAATCTGGAGCCGGTGAGCGTGGGTCTCGCGGTATCATTGCAGCACTGGGGCCAGATG

GTAAGCCCTCCCGTATCGTAGTTATCTACACGACGGGGAGTCAGGCAACTATGGATGAAC

GAAATAGACAGATCGCTGAGATAGGTGCCTCACTGATTAAGCATTGGTAACTGTCAGACC

AAGTTTACTCATATATACTTTAGATTGATTTAAAACTTCATTTTTAATTTAAAAGGATCT

AGGTGAAGATCCTTTTTGATAATCTCATGACCAAAATCCCTTAACGTGAGTTTTCGTTCC

ACTGAGCGTCAGACCCCGTAGAAAAGATCAAAGGATCTTCTTGAGATCCTTTTTTTCTGC

GCGTAATCTGCTGCTTGCAAACAAAAAAACCACCGCTACCAGCGGTGGTTTGTTTGCCGG

ATCAAGAGCTACCAACTCTTTTTCCGAAGGTAACTGGCTTCAGCAGAGCGCAGATACCAA

ATACTGTCCTTCTAGTGTAGCCGTAGTTAGGCCACCACTTCAAGAACTCTGTAGCACCGC

CTACATACCTCGCTCTGCTAATCCTGTTACCAGTGGCTGCTGCCAGTGGCGATAAGTCGT

GTCTTACCGGGTTGGACTCAAGACGATAGTTACCGGATAAGGCGCAGCGGTCGGGCTGAA

CGGGGGGTTCGTGCACACAGCCCAGCTTGGAGCGAACGACCTACACCGAACTGAGATACC

TACAGCGTGAGCTATGAGAAAGCGCCACGCTTCCCGAAGGGAGAAAGGCGGACAGGTATC

CGGTAAGCGGCAGGGTCGGAACAGGAGAGCGCACGAGGGAGCTTCCAGGGGGAAACGCCT

GGTATCTTTATAGTCCTGTCGGGTTTCGCCACCTCTGACTTGAGCGTCGATTTTTGTGAT

GCTCGTCAGGGGGGCGGAGCCTATGGAAAAACGCCAGCAACGCGGCCTTTTTACGGTTCC

TGGCCTTTTGCTGGCCTTTTGCTCACATGTTCTTTCCTGCGTTATCCCCTGATTCTGTGG

ATAACCGTATTACCGCCTTTGAGTGAGCTGATACCGCTCGCCGCAGCCGAACGACCGAGC

GCAGCGAGTCAGTGAGCGAGGAAGCGGAAGAGCGCCCAATACGCAAACCGCCTCTCCCCG

CGCGTTGGCCGATTCATTAATGCAGCTGGCACGACAGGTTTCCCGACTGGAAAGCGGGCA

GTGAGCGCAACGCAATTAATGTGAGTTAGCTCACTCATTAGGCACCCCAGGCTTTACACT

TTATGCTTCCGGCTCGTATGTTGTGTGGAATTGTGA

Restriction sites to linearize plasmid

Partial or complete disruption of the gene Partial

Additional remarks partial/complete disruption See for more information about the disruption Lasonder, E. et al., 2008, PloS Pathogens 10, e10000195 (Supporting Information: S3).

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 The genetic modification results in the disruption of the following gene models PB101363.00.0 and PB000829.02.0 and PB105739.00.0 (= PF14_0435).
See for more information about the disruption Lasonder, E. et al., 2008, PloS Pathogens 10, e10000195 (Supporting Information: S3).

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	AAAGGTACCGGTTTAATGTTGGTAGAC
Additional information primer 1	L2666 (Asp718); 5'
Sequence Primer 2	AAAAAGCTTGAGAATGGTTATAAGAC
Additional information primer 2	L2653 (HindIII); 5'
Sequence Primer 3	AAAGAATTCAGAAAATGCATTTTGAAG
Additional information primer 3	L2654 (EcoRI); 3'
Sequence Primer 4	AAAGCGGCCGCGATTATACAATACATATG
Additional information primer 4	L2655 (NotI); 3'
Sequence Primer 5
Additional information primer 5
Sequence Primer 6
Additional information primer 6

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