Back to search resultsSummaryRMgm-844
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Successful modification | The 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 Parasite | P. yoelii |
Parent strain/line | P. 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/Researcher | Pei Y; Kappe SHI |
Name Group/Department | Seattle Biomedical Research Institute, |
Name Institute | Seattle Biomedical Research Institute, |
City | Seattle |
Country | US |
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Details of the target gene | |||||||||||||||||||||||||
Gene Model of Rodent Parasite | PY17X_0816300 | ||||||||||||||||||||||||
Gene Model P. falciparum ortholog | PF3D7_0911900 | ||||||||||||||||||||||||
Gene product | falstatin | inhibitor of cysteine proteases | ||||||||||||||||||||||||
Gene product: Alternative name | ICP; falstatin | ||||||||||||||||||||||||
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Details of the genetic modification | |||||||||||||||||||||||||
Inducable system used | No | ||||||||||||||||||||||||
Additional remarks inducable system | |||||||||||||||||||||||||
Type of plasmid/construct used | (Linear) plasmid double cross-over | ||||||||||||||||||||||||
PlasmoGEM (Sanger) construct/vector used | No | ||||||||||||||||||||||||
Modified PlasmoGEM construct/vector used | No | ||||||||||||||||||||||||
Plasmid/construct map | |||||||||||||||||||||||||
Plasmid/construct sequence | |||||||||||||||||||||||||
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 | Multiple 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
Primer information: Primers used for amplification of the target sequences
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