SummaryRMgm-102
|
Successful modification | The parasite was generated by the genetic modification |
The mutant contains the following genetic modification(s) | Gene disruption |
Reference (PubMed-PMID number) |
Reference 1 (PMID number) : 16027235 |
MR4 number | |
top of page | |
Parent parasite used to introduce the genetic modification | |
Rodent Malaria Parasite | P. berghei |
Parent strain/line | P. berghei NK65 |
Name parent line/clone | Not applicable |
Other information parent line | |
top of page | |
The mutant parasite was generated by | |
Name PI/Researcher | A.S.I. Aly, K. Matuschewski |
Name Group/Department | Department of Parasitology |
Name Institute | Heidelberg University School of Medicine |
City | Heidelberg |
Country | Germany |
top of page | |
Name of the mutant parasite | |
RMgm number | RMgm-102 |
Principal name | ecp1(-) |
Alternative name | |
Standardized name | |
Is the mutant parasite cloned after genetic modification | Yes |
top of page | |
Phenotype | |
Asexual blood stage | Not different from wild type |
Gametocyte/Gamete | Not different from wild type |
Fertilization and ookinete | Not different from wild type |
Oocyst | Normal production of oocysts. Sporozoites fail to egress from midgut oocysts. Within the oocysts the sporozoites seemed to be arranged in circles whereas in wild type oocysts the sporozoites are arranged in the typical radial fashion. Mutant sporozoites displayed a continuous circular movement around a central axis, in both clockwise and anticlockwise directions. In wild type oocysts, sporozoite bending and flexing is seen on rare occasions and in general, no motility can be observed in wild type oocysts. Mutant oocysts were more resistant to permeabilization by detergents (1% saponin) compared to wild type oocysts. |
Sporozoite | Sporozoites fail to egress from midgut oocysts. No sporozoites were detected in the hemocoel or in the salivary glands of infected mosquitoes despite efficient infection rates and high numbers of oocyst sporozoites. Mechanically liberated sporozoites from oocysts show gliding motility. Infection of Sprague/Dawley rats by intravenous injection of large numbers of mechanically liberated sporozoites (100.000) did not result in blood stage infections. |
Liver stage | Infection of Sprague/Dawley rats by intravenous injection of large numbers of mechanically liberated sporozoites (100.000) did not result in blood stage infections. |
Additional remarks phenotype | Mutant/mutation Plasmodium species possesses SERAs of two major groups, specified as 'cysteine-type SERAs' and 'serine-type SERAs'. Serine-type SERAs contain an active site serine residue instead of the canonical cysteine residue. The genomes of different Plasmodium species contain different numbers of SERA genes. P. falciparum possesses nine SERA protease genes whereas the P. berghei genome contains five. P. falciparum possesses six “serine-type” (SERA1 to SERA5 and SERA9) and three “cysteine-type” (SERA6 to SERA8) SERAs. In P. falciparum mutants have been generated lacking expression of SERA1, 2, 3, 4 , 7, 8 and 9 without a distinct phenotype in the blood stages (MvCoubrie J.E. et al, 2007, Infection and Immunity, 75:5565-74). SERA genes of P. berghei are arranged in a tandem cluster that contains two serine-type SERAs (PbSERA1, -2) and three cysteine-type SERAs (PbSERA3, -4 and -5). Sequence homology and syntenic organisation indicate that PbSERA3, -4 (PB000352.01.0) and -5 (PB000649.01.0) are orthologs of P. falciparum SERA6, -7, -8. Phenotype Additional information Other mutants Other mutants
|
top of page | |||||||||||||||||||||||||
Details of the target gene | |||||||||||||||||||||||||
Gene Model of Rodent Parasite | PBANKA_0304700 | ||||||||||||||||||||||||
Gene Model P. falciparum ortholog | PF3D7_0207300 | ||||||||||||||||||||||||
Gene product | serine repeat antigen 8 | ||||||||||||||||||||||||
Gene product: Alternative name | ECP1, egress cysteine protease 1; PbSERA5 | ||||||||||||||||||||||||
top of page | |||||||||||||||||||||||||
Details of the genetic modification | |||||||||||||||||||||||||
Inducable system used | No | ||||||||||||||||||||||||
Additional remarks inducable system | |||||||||||||||||||||||||
Type of plasmid/construct used | Plasmid single 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 | Partial | ||||||||||||||||||||||||
Additional remarks partial/complete disruption | The ecp1 genomic locus is targeted with an EcoRV-linearized integration plasmid containing 5' and 3' truncations of the ecp1 open reading frame and the dhfr/ts positive selectable marker. Upon a single crossover event, the region of homology is duplicated, resulting in two truncated, non expressed ecp1 copies in the recombinant locus. | ||||||||||||||||||||||||
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 | |||||||||||||||||||||||||
Additional remarks selection procedure | |||||||||||||||||||||||||
Primer information: Primers used for amplification of the target sequences
![]() Primer information: Primers used for amplification of the target sequences
![]()
| |||||||||||||||||||||||||
top of page |