SummaryRMgm-1582
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Successful modification | The parasite was generated by the genetic modification |
The mutant contains the following genetic modification(s) | Gene disruption, Introduction of a transgene |
Reference (PubMed-PMID number) |
Reference 1 (PMID number) : 27600503 |
MR4 number | |
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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 | Not applicable |
Other information parent line | |
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The mutant parasite was generated by | |
Name PI/Researcher | Rizopoulos Z, Matuschewski K, Haussig J. |
Name Group/Department | Parasitology Unit |
Name Institute | Max Planck Institute for Infection Biology |
City | Berlin |
Country | Germany |
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Name of the mutant parasite | |
RMgm number | RMgm-1582 |
Principal name | alas(-) |
Alternative name | |
Standardized name | |
Is the mutant parasite cloned after genetic modification | Yes |
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Phenotype | |
Asexual blood stage | Not different from wild type |
Gametocyte/Gamete | Not tested |
Fertilization and ookinete | Not different from wild type |
Oocyst | Normal numbers of oocysts are formed. However, day 10 and 14 oocysts had a reduced size (`60%) compared to wild type oocysts. A large (~10-20-fold) reduction in the numbers of midgut-associated sporozoites was observed. No hemocoel or salivary gland sporozoites were detected. |
Sporozoite | Normal numbers of oocysts are formed. However, day 10 and 14 oocysts had a reduced size (`60%) compared to wild type oocysts. A large (~10-20-fold) reduction in the numbers of midgut-associated sporozoites was observed. No hemocoel or salivary gland sporozoites were detected. |
Liver stage | A large (~10-20-fold) reduction in the numbers of midgut-associated sporozoites was observed. No hemocoel or salivary gland sporozoites were detected. No mosquito transmission. See also additional information of the role of ALAS during liver stage development |
Additional remarks phenotype | Mutant/mutation Phenotype Additional information Some results in this paper are different from other analyses of mutants lacking expression of ALAS. See mutant RMgm-924 that showed strongly reduced oocyst production and absence of liver stage development. |
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Details of the target gene | |||||||||||||||||||||||||
Gene Model of Rodent Parasite | PBANKA_1459200 | ||||||||||||||||||||||||
Gene Model P. falciparum ortholog | PF3D7_1246100 | ||||||||||||||||||||||||
Gene product | delta-aminolevulinic acid synthetase | ||||||||||||||||||||||||
Gene product: Alternative name | ALAS | ||||||||||||||||||||||||
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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 | hdhfr/yfcu | ||||||||||||||||||||||||
Promoter of the selectable marker | pbdhfr | ||||||||||||||||||||||||
Selection (positive) procedure | pyrimethamine | ||||||||||||||||||||||||
Selection (negative) procedure | No | ||||||||||||||||||||||||
Additional remarks genetic modification | Fragments of the 3’ and 5’ UTR were amplified from gDNA using gene-specific primers. First, 3’ fragments were cloned into the pBAT vector, following restriction digestion with HindIII and KpnI to generate an intermediate construct (pHEME-im). Then, the 5’ UTR fragment was digested with SacII and EcoRV and cloned into the respective SacII- and PvuII-linearized pHEME-im construct, thus removing the mCherry-3xMyc tag. Finally, the pHEME-ko vector was linearized with ScaI and SalI before transfection into P. berghei strain ANKA (wt) parasites. | ||||||||||||||||||||||||
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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Type and details of transgene | |||||||||||||||||||
Is the transgene Plasmodium derived | Transgene: not Plasmodium | ||||||||||||||||||
Transgene name | GFP | ||||||||||||||||||
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Details of the genetic modification | |||||||||||||||||||
Inducable system used | No | ||||||||||||||||||
Additional remarks inducable system | |||||||||||||||||||
Type of plasmid/construct | (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 | |||||||||||||||||||
Selectable marker used to select the mutant parasite | hdhfr/yfcu | ||||||||||||||||||
Promoter of the selectable marker | pbdhfr | ||||||||||||||||||
Selection (positive) procedure | pyrimethamine | ||||||||||||||||||
Selection (negative) procedure | No | ||||||||||||||||||
Additional remarks genetic modification | |||||||||||||||||||
Additional remarks selection procedure | |||||||||||||||||||
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Other details transgene | |||||||||||||||||||
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Promoter | |||||||||||||||||||
Gene Model of Parasite | PBANKA_0711900 | ||||||||||||||||||
Gene Model P. falciparum ortholog | PF3D7_0818900 | ||||||||||||||||||
Gene product | heat shock protein 70 | ||||||||||||||||||
Gene product: Alternative name | HSP70 | ||||||||||||||||||
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3'-UTR | |||||||||||||||||||
Gene Model of Parasite | PBANKA_1340000 | ||||||||||||||||||
Gene product | dihydrofolate synthase/folylpolyglutamate synthase, putative | ||||||||||||||||||
Gene product: Alternative name | PbDHFS-FPGS | ||||||||||||||||||
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Insertion/Replacement locus | |||||||||||||||||||
Replacement / Insertion | Replacement locus | ||||||||||||||||||
Gene Model of Parasite | PBANKA_1459200 | ||||||||||||||||||
Gene product | delta-aminolevulinic acid synthetase | ||||||||||||||||||
Gene product: Alternative name | ALAS | ||||||||||||||||||
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