Additional remarks phenotype | Mutant/mutation
The mutant lacks expression of PBANKA_0519500.
Protein (function)
Phenotype
Reduced growth/multiplication of asexual blood stages.
Normal gametocyte production, male gamete formation (exflagellation), oocyst and sporozoite formation. Sporozoite motility and speed reduced.
Additional information
In independent transfection experiments 50 genes were targeted genes for deletion (using PlasmoGem gene-deletion DNA constructs). For 26 genes, populations of mixed wild type and transgenic parasites were selected. From those populations transgenic clonal lines for 14 genes could be isolated (see below).
Around half of these 14 clonal mutant parasite lines showed reduced blood stage growth rates comparable to those observed in the previously published PlasmoGEM screen with narrow confidence intervals of PlasmoGEM growth rates being a predictor for the growth rate of clonal lines. The other half showed growth rates closer to the growth rate of wild type parasites.
Life cycle progression of the 14 gene deletion mutants through mosquitoes was analysed, including 3 existing gene deletion mutants with reduced growth/multiplication of asexual blood stages: Plasmepsin IV (-) (mutant RMgm-314; PBANKA_1034400; PF3D7_1408100); Aminopeptidase P (-) (mutant RMgm-813; PBANKA_1318100; PF3D7_1454400); l-aminopeptidase (-) (mutant RMgm-814; PBANKA_1309900; PF3D7_1446200).
Eight of the 17 mutants mutants showed defects before sporozoite formation. These mutants lacked the genes encoding plasmepsin IV (no/block sporozoite formation), profilin (no/block oocyst formation), beta-catenin like protein 1 (no male gamete formation/exflagellation), telomeric DNA-binding protein (no/block oocyst formation), mannose-1-phosphate guanyltransferase (man-1-P GT)(no/block oocyst formation), U2 snRNP-associated SURP motif-containing protein (U2snRNP)(no male gamete formation/exflagellation), diacylglycerol kinase (no salivary gland sporozoites) and hypoxanthine-guanine phosphoribosyl transferase (hypox-guan phosph transf) (no/block gametocyte formation).
The other nine mutant lines were able to complete the life cycle. These mutants lacked the genes encoding dynein heavy chain, autophagy-related protein 23 (autophagy-rel protein 23), tubulin tyrosine ligase, a conserved protein, dipeptidyl aminopeptidase 1 (dipeptidyl aminopept 1), protein kinase, V-type(+) pyrophosphatase (V-type(+) pyrophosph), aminopeptidase P (app) or M17 leucyl aminopeptidase (lap).
For 5 out of the 17 mutants infection of C57Bl6 mice led to a delayed infection that could be controlled and cleared by the mice. These 5 mutants lacked the genes encoding aminopeptidase P (app) or M17 leucyl aminopeptidase (lap), U2 snRNP-associated SURP motif-containing protein (U2snRNP), diacylglycerol kinase and hypoxanthine-guanine phosphoribosyl transferase (hypox-guan phosph transf).
Dynein heavy chain: PBANKA_0615700, PF3D7_0718000
Autophagy-related protein 23: PBANKA_0921700, PF3D7_1126700
Profilin: PBANKA_0833000, PF3D7_0932200
Tubulin tyrosine ligase: PBANKA_0901900, PF3D7_1147200
Telomeric DNA binding protein: PBANKA_1205000, PF3D7_1006800
Beta-catenin like protein: PBANKA_0910100, PF3D7_1138600
Man-1-P-GT: PBANKA_1022300, PF3D7_1420900
Diacylglycerol kinase: PBANKA_1334600, PF3D7_1471400
U2 snRNP: PBANKA_1039300 PF3D7_1402700
Conserved protein: PBANKA_0519500, No
Dipeptidyl aminopeptidase: PBANKA_0931300, PF3D7_1116700
Hypoxanthine-guanine phosphoribosyl transferase: PBANKA_1210800, PF3D7_1012400
Protein kinase: PBANKA_0826900, PF3D7_0926100
V-type H(+)-translocating pyrophosphatase: PBANKA_1320500, PF3D7_1235200
Other mutants |