RMgmDB - Rodent Malaria genetically modified Parasites

Summary

RMgm-5213

Malaria parasite	P. berghei
Genotype
Disrupted	Gene model (rodent): PBANKA_1457700; Gene model (P.falciparum): PF3D7_1244500; Gene product: PIMMS57 protein (PSOP26)
Phenotype	Fertilization and ookinete; Oocyst;

Last modified: 20 June 2022, 14:27

*RMgm-5213

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) : 35606790
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	P. berghei ANKA 2.34
Other information parent line	P. berghei ANKA 2.34 is a cloned, gametocyte producer line of the ANKA strain (PubMed: PMID: 15137943).
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The mutant parasite was generated by
Name PI/Researcher	Wang PP, Cao Y
Name Group/Department	Department of Immunology, College of Basic Medical Sciences
Name Institute	China Medical University
City	Shenyang
Country	China
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Name of the mutant parasite
RMgm number	RMgm-5213
Principal name	Δpsop26
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 different from wild type
Fertilization and ookinete	A strong reduction (>69%) in mature ookinete numbers.
Oocyst	Reduced oocyst numbers (80% reduction).
Sporozoite	Not tested
Liver stage	Not tested
Additional remarks phenotype	Mutant/mutation The mutant lacks expression of PSOP26 (PIMMS57 protein) Protein (function) See also mutant RMgm-4998 Phenotype A strong reduction (>69%) in mature ookinete numbers. Reduced oocyst numbers (80% reduction). 'The zygote numbers in Δpsop26 at 2 h after incubating at 19 °C were comparable to wild-type, indicating that the reduced matured ookinete number in Δpsop26 mutants is caused by an impairment of the zygote-to-ookinete developmental transition.' See also mutant RMgm-4998 lacking expression of PSOP26/PIMMS57 protein where they found a (slightly) different phenotype ('wild type-like ookinete production but reduced oocyst numbers'). Additional information Analysis of a mutant expressing a C-terminal 3xHA tagged version of PSOP26 (RMgm-5214) provided evidence for localization of the protein at the membrane of male and female gametes (absent in gametocytes before activation) and at the membrane of ookinetes. See also mutant RMgm-5001 where the found evidence for expression in (the membrane of) ookinetes of PSOP26/PIMMS57 Other mutants

Disrupted: Mutant parasite with a disrupted gene

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Details of the target gene

Gene Model of Rodent Parasite

PBANKA_1457700

Gene Model P. falciparum ortholog

PF3D7_1244500

Gene product

PIMMS57 protein

Gene product: Alternative name

PSOP26

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Details of the genetic modification

Inducable system used

Additional remarks inducable system

Type of plasmid/construct used

(Linear) plasmid double cross-over

PlasmoGEM (Sanger) construct/vector used

Yes

Name of PlasmoGEM construct/vector

PbGEM-325763

Modified PlasmoGEM construct/vector used

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

eef1a

Selection (positive) procedure

pyrimethamine

Selection (negative) procedure

Additional remarks genetic modification

To disrupt the psop26 gene, a ∆psop26 transgenic parasite was generated using the PbGEM325763 plasmid

Additional remarks selection procedure

Primer information: Primers used for amplification of the target sequences Click to view information

Primer information: Primers used for amplification of the target sequences Click to hide information

Sequence Primer 1
Additional information primer 1
Sequence Primer 2
Additional information primer 2
Sequence Primer 3
Additional information primer 3
Sequence Primer 4
Additional information primer 4
Sequence Primer 5
Additional information primer 5
Sequence Primer 6
Additional information primer 6

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