top of page |
Details of the target gene |
Gene Model of Rodent Parasite |
PBANKA_0931200
|
Gene Model P. falciparum ortholog |
PF3D7_1116800
|
Gene product | heat shock protein 101 | chaperone protein ClpB2 |
Gene product: Alternative name | HSP101 |
top of page |
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 | |
Additional remarks selection procedure | The unsuccessful attempts to disrupt the hsp101 gene indicates an essential role of HSP101 in blood stages.
Plasmodium parasites remodel their vertebrate host cells by translocating hundreds of proteins across an encasing membrane into the host cell cytosol via a putative export machinery termed PTEX (Plasmodium Translocon of EXported protein). HSP101 (PbANKA_094120), PTEX150 (PbANKA_100850), EXP2 (PbANKA_133430), PTEX88 (PbANKA_094130) and TRX2 (PbANKA_135800) have been identified as members of the PTEX complex.
These proteins are also expressed in early gametocytes, mosquito and liver stages. Although amenable to genetic tagging, HSP101, PTEX150, EXP2 and PTEX88 could not be genetically deleted in P. berghei, in keeping with the obligatory role this complex is postulated to have in maintaining normal blood-stage growth. In contrast, the putative thioredoxin-like protein TRX2 could be deleted, with knockout parasites displaying reduced grow-rates, both in vivo and in vitro. |
Primer information: Primers used for amplification of the target sequences
Primer information: Primers used for amplification of the target sequences
Sequence Primer 1 | tagggcccGCTCTTAATGCATTTGCTTTATA |
Additional information primer 1 | S23F: PbHSP101 5’ UTR (ApaI) |
Sequence Primer 2 | gttccgcggGCAATGTTCCGTACCATTTCTACT |
Additional information primer 2 | S24F: PbHSP101 5’ UTR (SacII) |
Sequence Primer 3 | gatatcGGCTCATTATCATTTGAATCATCA |
Additional information primer 3 | S25bF: PbHSP101 3’ UTR (EcoRV) |
Sequence Primer 4 | gcggccgcGTGTTGGTCTAGCTCCTAATTCTG |
Additional information primer 4 | S25bR: PbHSP101 3’ UTR (NotI) |
Sequence Primer 5 | |
Additional information primer 5 | |
Sequence Primer 6 | |
Additional information primer 6 | |
|
top of page |