RMgmDB - Rodent Malaria genetically modified Parasites
SummaryRMgm-64
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Last modified: 27 July 2015, 18:22
*RMgm-64| 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 | 2 |
| Reference (PubMed-PMID number) | Not published (yet) |
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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 | 8417HP |
| Other information parent line | A reference wild type clone from the ANKA strain of P. berghei (Janse et al., Exp. Parasitol. 68, 274-282). |
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| Attempts to generate the mutant parasite were performed by | |
| Name PI/Researcher | Pace, T; Ponzi, M |
| Name Group/Department | Laboratorio di Biologia Cellulare |
| Name Institute | Istituto Superiore di Sanita |
| City | Rome |
| Country | Italy |
Disrupted: Mutant parasite with a disrupted gene| top of page | |||||||||||||||||||||||||
| Details of the target gene | |||||||||||||||||||||||||
| Gene Model of Rodent Parasite | PBANKA_0524200 | ||||||||||||||||||||||||
| Gene Model P. falciparum ortholog | Not available | ||||||||||||||||||||||||
| Gene product | early transcribed membrane protein; small exported protein | ||||||||||||||||||||||||
| Gene product: Alternative name | SEP2; ETRAMP4; Pbsep2 | ||||||||||||||||||||||||
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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 | pbdhfr | ||||||||||||||||||||||||
| Promoter of the selectable marker | pbdhfr | ||||||||||||||||||||||||
| Selection (positive) procedure | pyrimethamine | ||||||||||||||||||||||||
| Selection (negative) procedure | No | ||||||||||||||||||||||||
| Additional remarks genetic modification | In P. berghei, this gene belongs to a family of three related genes (encoding so-called SEP proteins; sep1-3), located in the subtelomeric regions of chromosomes 5 (Birago et al., Mol. Biochem. Parasitol. (2003) 126, 209-18). The SEP proteins show homology to members of the ETRAMP family of proteins of P. falciparum but the orthologous relationship of the different members is unclear (ortholog of etramp4, PFD1120c?). The gene encodes encodes an integral membrane protein. The protein contains a predicted signal peptide at the NH(2)-terminus, an internal hydrophobic region and a polymorphic, low-complexity region at the carboxy-terminus (Birago et al., Mol. Biochem. Parasitol. (2003) 126, 209-18). The failure to disrupt sep2 (sep-bis) has not been published yet. Two transfections has been performed using a BioRad electroporation device(pers. comm. M. Ponzi, T. Pace). The coding region of this gene as confirmed by ESTs is as follows (pers. comm. Dr. T. Pace): MKLAKALYFVAFLLAIKVLTPGSNNYVEAKPANSKKVTKSGDNAFIKKIKNNKAAFISTLAA TVALAIATTFGVMHYQNNGGFEKSPWANKIDKNKSLPKKKTDNSPPSKGNLGNTSQKRNPISF TPISDGYKPSSSPNNNSKRTPTAPYTVKLN The two other sep genes, sep1 (PB100384.00.0; PF11_0039; early transcribed membrane protein, etramp) and sep3 (PB401867.00.0; sep-ter) have been targeted for gene disruption. See RMgm-16 and RMgm-18 for mutants containing targeted disruptions of sep1 See RMgm-65 for more information on sep3. | ||||||||||||||||||||||||
| Additional remarks selection procedure | |||||||||||||||||||||||||
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Primer information: Primers used for amplification of the target sequences
 Primer information: Primers used for amplification of the target sequences
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