RMgmDB - Rodent Malaria genetically modified Parasites

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Summary

RMgm-4707

Malaria parasite	P. berghei
Genotype
Disrupted	Gene model (rodent): PBANKA_1205000; Gene model (P.falciparum): PF3D7_1006800; Gene product: single-strand telomeric DNA-binding protein GBP2, putative (GBP2, G-strand binding protein 2)
Phenotype	Asexual bloodstage; Gametocyte/Gamete;

Last modified: 21 January 2020, 17:48

*RMgm-4707

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) : 31958569
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	Niikura M, Kobayashi F
Name Group/Department	Department of Environmental Science, School of Life and Environmental Science
Name Institute	Azabu University
City	-
Country	Japan
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Name of the mutant parasite
RMgm number	RMgm-4707
Principal name	Δgbp2
Alternative name
Standardized name
Is the mutant parasite cloned after genetic modification	Yes
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Phenotype
Asexual blood stage	Reduced growth/multiplication of asexual blood stages. Blood stage infections induce experimental cerebral malaria (ECM) in C57Bl6 mice.
Gametocyte/Gamete	(Strongly) reduced numbers of male and female gametocytes
Fertilization and ookinete	Not tested
Oocyst	Not tested
Sporozoite	Not tested
Liver stage	Not tested
Additional remarks phenotype	Mutant/mutation The mutant lacks expression of GBP2 Protein (function) G-strand binding protein 2 (GBP2) is a Ser/Arg-rich (SR) protein involved in mRNA surveillance and nuclear mRNA quality control in yeast. PfGBP2 (PF3D7_1006800) has homology to yeast GBP2 and is highly expressed during the blood stages of malaria parasites. GBP2 of Plasmodium spp. contains two RNA recognition motif (RRM) domains (RRM1 and RRM2) and is conserved among eukaryotes. RRM1 and RRM2 of yeast GBP2 recognize RNA. RRM2 of yeast GBP2 contains a WQxLKD motif, which is directly involved in RNA recognition. Plasmodium RRM2 contains a WKELKD motif, which is similar to the WQxLKD motif. In contrast, RRM3 (the C-terminal RRM domain of yeast) was absent in Plasmodium spp. Phenotype Reduced growth/multiplication of asexual blood stages. Blood stage infections induce experimental cerebral malaria (ECM) in C57Bl6 mice. (Strongly) reduced numbers of male and female gametocytes. Additional information Other mutants

Disrupted: Mutant parasite with a disrupted gene

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Details of the target gene

Gene Model of Rodent Parasite

PBANKA_1205000

Gene Model P. falciparum ortholog

PF3D7_1006800

Gene product

single-strand telomeric DNA-binding protein GBP2, putative

Gene product: Alternative name

GBP2, G-strand binding protein 2

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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

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

Promoter of the selectable marker

unknown

Selection (positive) procedure

pyrimethamine

Selection (negative) procedure

Additional remarks genetic modification

The gene-targeting vector for GBP2, gbp2 (PBANKA_120500), was prepared by PCR. Briefly, the 5'and 3' flanking regions of the gbp2 open reading frame (ORF) were amplified by PCR. The PCR products were annealed to either side of the red fluorescent protein gene (mCherry)-hdhfr-expressing cassette and amplified by PCR using gene-specific primers

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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