RMgmDB - Rodent Malaria genetically modified Parasites

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Summary

RMgm-844
Malaria parasiteP. yoelii
Genotype
Genetic modification not successful
DisruptedGene model (rodent): PY17X_0816300; Gene model (P.falciparum): PF3D7_0911900; Gene product: falstatin | inhibitor of cysteine proteases (ICP; falstatin)
PhenotypeNo phenotype has been described
Last modified: 26 September 2016, 12:09
  *RMgm-844
Successful modificationThe gene/parasite could not be changed/generated by the genetic modification.
The following genetic modifications were attempted Gene disruption
Number of attempts to introduce the genetic modification Unknown
Reference (PubMed-PMID number) Reference 1 (PMID number) : 23421981
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Parent parasite used to introduce the genetic modification
Rodent Malaria ParasiteP. yoelii
Parent strain/lineP. y. yoelii 17XNL
Name parent line/clone Not applicable
Other information parent line
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Attempts to generate the mutant parasite were performed by
Name PI/ResearcherPei Y; Kappe SHI
Name Group/DepartmentSeattle Biomedical Research Institute,
Name InstituteSeattle Biomedical Research Institute,
CitySeattle
CountryUS
  Disrupted: Mutant parasite with a disrupted gene
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Details of the target gene
Gene Model of Rodent Parasite PY17X_0816300
Gene Model P. falciparum ortholog PF3D7_0911900
Gene productfalstatin | inhibitor of cysteine proteases
Gene product: Alternative nameICP; falstatin
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Details of the genetic modification
Inducable system usedNo
Additional remarks inducable system
Type of plasmid/construct used(Linear) plasmid 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 modificationMultiple unsuccessful attempts to disrupt the icp gene (using both single and double cross over strategies) indicates that ICP is essential for blood stage growth.

HOWEVER: See RMgm-970 for successful disruption of the P. berghei icp gene!!!

See RMgm-845 for a mutant expressing a normal ICP and a quadruple C-myc tagged version of ICP.

ICP is conserved among numerous Plasmodium species. However, a Py-ICP ortholog had not been annotated in PlasmoDB (www.plasmodb.org). To determine if Py contained an ICP ortholog, a BLAST search of the Py genome was conducted using Pf-ICP as the query and observed highly conserved nucleotide sequences at the C-terminal region of a 7.3 kb gene, PY03424. It was found that that PY03424 was composed of two separate genes: a single exon gene is termed PY03424* and Py-ICP. The re-annotated Py-ICP gene has 85% and 34% amino acid identity to its orthologs in Pb and Pf, respectively.
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
Additional information primer 1
Sequence Primer 2
Additional information primer 2
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
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Conceptual design of the database: Dr. C.J. Janse (Leiden Malaria Research Group, Leiden, The Netherlands).
Technical design and realization: StudioDrie StudioDrie; S. Mededovic and A. van Wigcheren