RMgmDB - Rodent Malaria genetically modified Parasites

Summary

RMgm-88
Malaria parasiteP. berghei
Genotype
DisruptedGene model (rodent): PBANKA_0619200; Gene model (P.falciparum): PF3D7_0721700; Gene product: secreted ookinete protein, putative (PSOP1, putative secreted ookinete protein 1)
PhenotypeNo phenotype has been described
Last modified: 19 February 2009, 20:55
  *RMgm-88
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) : 18761621
MR4 number
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Parent parasite used to introduce the genetic modification
Rodent Malaria ParasiteP. berghei
Parent strain/lineP. berghei ANKA
Name parent line/clone P. berghei ANKA 2.34
Other information parent lineP. berghei ANKA 2.34 is a cloned, gametocyte producer line of the ANKA strain (PubMed: PMID: 15137943).
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The mutant parasite was generated by
Name PI/ResearcherA. Ecker; R.E. Sinden
Name Group/DepartmentDivision of Cell and Molecular Biology
Name InstituteImperial College
CityLondon
CountryUnited Kingdom
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Name of the mutant parasite
RMgm numberRMgm-88
Principal nameΔpsop1
Alternative name
Standardized name
Is the mutant parasite cloned after genetic modificationYes
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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 PSOP1 (putative secreted ookinete protein).

Protein (function)
Unknown. The protein is detected in a proteome analysis of ookinetes and contains a predicted signal sequence.

Phenotype
The lack of expression of PSOP1 does not result in a clear phenotype. In this study no in depht analysis of the phenotype of the different life cycle stages has been performed. Numbers salivary gland sporozoites in Anopheles stephensi are normal. Oocyst production is variable and lower compared to wildtype (but not significantly different). The mutant could 'easily be transmitted' by A. stephensi.

  Disrupted: Mutant parasite with a disrupted gene
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Details of the target gene
Gene Model of Rodent Parasite PBANKA_0619200
Gene Model P. falciparum ortholog PF3D7_0721700
Gene productsecreted ookinete protein, putative
Gene product: Alternative namePSOP1, putative secreted ookinete protein 1
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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
Plasmid/construct sequence
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 1TTGGGCCCACATATACATGTATGTACACCTG
Additional information primer 1AE01a (ApaI); 5'
Sequence Primer 2CCAAGCTTCTGTCTTATTAAGATTCGAGATG
Additional information primer 2AE01b (HindIII); 5'
Sequence Primer 3TGAATTCTTGCACAGGATGACATAGATGGC
Additional information primer 3AE01c (EcoRI); 3'
Sequence Primer 4GGGGATCCCATACCTACTCCCATGTGTGCAC
Additional information primer 4AE01d (BamHI); 3'
Sequence Primer 5
Additional information primer 5
Sequence Primer 6
Additional information primer 6
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Conceptual design of the database: Dr. C.J. Janse (Leiden Malaria Research Group, Leiden, The Netherlands).
Technical design and realization: S. Mededovic and A. van Wigcheren