Back to search resultsSummaryRMgm-963
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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 | ≥ 5 |
Reference (PubMed-PMID number) |
Reference 1 (PMID number) : 24244852 |
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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 2.34 |
Other information parent line | P. berghei ANKA 2.34 is a cloned, gametocyte producer line of the ANKA strain (PubMed: PMID: 15137943). |
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Attempts to generate the mutant parasite were performed by | |
Name PI/Researcher | B. Poulin; R. Tewari |
Name Group/Department | Centre for Genetics and Genomics |
Name Institute | School of Life Sciences, Queens Medical Centre, University of Nottingham |
City | Nottingham |
Country | UK |
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Details of the target gene | |||||||||||||||||||||||||
Gene Model of Rodent Parasite | PBANKA_1209400 | ||||||||||||||||||||||||
Gene Model P. falciparum ortholog | PF3D7_1011000 | ||||||||||||||||||||||||
Gene product | inner membrane complex sub-compartment protein 1 | ||||||||||||||||||||||||
Gene product: Alternative name | ISP1 | ||||||||||||||||||||||||
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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 | |||||||||||||||||||||||||
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 | Attempts to knock-out isp1 were unsuccessful indicating an essential role of ISP1 for asexual blood stage growth/multiplication. Four IMC sub-compartment proteins (ISPs) were described in T. gondii coded by genes restricted to the Apicomplexa. ISPs are small proteins of approximately 150 amino acids and usually characterized by a MetGly(Xaa)2–5CysCys sequence motif at the N-terminus (except for ISP4), but are otherwise relatively divergent and without either obvious domains, low complexity sequence or homology to other known proteins. To identify ISPs in Plasmodium, an ISP-specific hidden Markov model was constructed from the four ISPs previously identified in T. gondii. Plasmodium species were shown to contain only genes for ISP1 (PF3D7_1011000) and ISP3 (PF3D7_1460600). These families are the most conserved in terms of protein sequence and domain architecture and generally have strong predictions for N-myristoylation and palmitoylation. RMgm-962: A mutant lacking expression of inner membrane complex sub-compartment protein 3, putative (ISP3; PBANKA_132430) RMgm-964: A mutant expressing a C-terminal GFP tagged version of ISP3 RMgm-965: A mutant expressing a C-terminal GFP tagged version of ISP1 (PBANKA_120940) | ||||||||||||||||||||||||
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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