RMgmDB - Rodent Malaria genetically modified Parasites

Summary

RMgm-1300

Malaria parasite	P. berghei
Genotype
Tagged	Gene model (rodent): PBANKA_1119200; Gene model (P.falciparum): PF3D7_0620000; Gene product: secreted ookinete protein 25, putative \| ookinete surface-associated protein 8, putative (POS8; putative ookinete surface-associated protein 8; PSOP25) Name tag: GFP
Phenotype	No phenotype has been described

Last modified: 7 November 2023, 17:50

*RMgm-1300

Successful modification	The parasite was generated by the genetic modification
The mutant contains the following genetic modification(s)	Gene tagging
Reference (PubMed-PMID number)	Reference 1 (PMID number) : 26018192
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	Kaneko I; Yuda M
Name Group/Department	Department of Medical Zoology
Name Institute	Mie University Graduate School of Medicine
City	Tsu, Mie
Country	Japan
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Name of the mutant parasite
RMgm number	RMgm-1300
Principal name	POS8-GFP
Alternative name
Standardized name
Is the mutant parasite cloned after genetic modification	Yes
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Phenotype
Asexual blood stage	Not tested
Gametocyte/Gamete	Not tested
Fertilization and ookinete	Not different from wild type
Oocyst	Not tested
Sporozoite	Not tested
Liver stage	Not tested
Additional remarks phenotype	Mutant/mutation The mutant expressed a GFP-tagged version of POS8 (putative ookinete surface-associated protein. 8). The tagged gene is introduced on the centromere plasmid pCen-GFP (circular PAC). Protein (function) The protein was identified in a genome-wide screen of target genes of AP2-O by chromatin immunoprecipitation-sequencing. Phenotype No GFP signals could be detected in ookinetes Phenotype analysis of a mutant lacking expression of POS8 (RMgm-1299) showed reduced oocyst production (and reduced size of oocysts) Additional information Other mutants RMgm-1299: A mutant lacking expression of POS8

Tagged: Mutant parasite with a tagged gene

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Details of the target gene

Gene Model of Rodent Parasite

PBANKA_1119200

Gene Model P. falciparum ortholog

PF3D7_0620000

Gene product

secreted ookinete protein 25, putative | ookinete surface-associated protein 8, putative

Gene product: Alternative name

POS8; putative ookinete surface-associated protein 8; PSOP25

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Details of the genetic modification

Name of the tag

GFP

Details of tagging

C-terminal

Additional remarks: tagging

To examine localisation of the protein, the ORF was expressed as a GFP-tagged protein under control of the original promoter, using the centromere plasmid pCen-GFP.

Commercial source of tag-antibodies

Type of plasmid/construct

Plasmodium Artificial Chromosome (PAC) circulair

PlasmoGEM (Sanger) construct/vector used

Modified PlasmoGEM construct/vector used

Plasmid/construct map

Plasmid/construct sequence

Restriction sites to linearize plasmid

Selectable marker used to select the mutant parasite

tgdhfr

Promoter of the selectable marker

eef1a

Selection (positive) procedure

pyrimethamine

Selection (negative) procedure

Additional remarks genetic modification

Protein targeting was investigated by expressing the gene as a GFP-tagged protein, using the centromere plasmid pCen-GFP (circular PAC). Target genes with the upstream regulatory region (1.1–1.5-kbp upstream of the first methionine codon) were amplified by PCR using genomic DNA as the template, and then subcloned into the region immediately upstream of the GFP gene of the pCen vector.

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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