RMgmDB - Rodent Malaria genetically modified Parasites

Summary

RMgm-5087

Malaria parasite	P. berghei
Genotype
Disrupted	Gene model (rodent): PBANKA_1230500; Gene model (P.falciparum): PF3D7_0615700; Gene product: THO complex subunit 4, putative (THO4)
Phenotype	Asexual bloodstage; Gametocyte/Gamete;

Last modified: 26 October 2021, 15:09

*RMgm-5087

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) : 34604117
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	Kanagawa
Country	Japan
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Name of the mutant parasite
RMgm number	RMgm-5087
Principal name	Δtho4
Alternative name
Standardized name
Is the mutant parasite cloned after genetic modification	Yes
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Phenotype
Asexual blood stage	Normal (wild type) growth of asexual blood stages.
Gametocyte/Gamete	The percentage of male gametocytes in cultured Δtho4 parasites was comparable to the percentage in control parasites. The percentage of female gametocytes was lower in cultured Δtho4 parasites than in control parasites.
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 THO4 Protein (function) Nuclear poly(A) binding protein 2 (NAB2), THO complex subunit 4 (THO4), nucleolar protein 3 (NPL3), G-strand binding protein 2 (GBP2) and serine/arginine-rich splicing factor 1 (SR1) are involved in nuclear mRNA export in malaria parasites. NTHO complex subunit 4 (THO4) is a homologue of yeast YRA1 in malaria parasites. THO4 is characterized by its central RNA-binding domain. Phenotype Normal (wild type) growth of asexual blood stages. The percentage of male gametocytes in cultured Δtho4 parasites was comparable to the percentage in control parasites. The percentage of female gametocytes was lower in cultured Δtho4 parasites than in control parasites. 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_1230500

Gene Model P. falciparum ortholog

PF3D7_0615700

Gene product

THO complex subunit 4, putative

Gene product: Alternative name

THO4

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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

eef1a

Selection (positive) procedure

pyrimethamine

Selection (negative) procedure

Additional remarks genetic modification

To generated nab2-, tho4-, npl3- and sr1-deleted P. berghei ANKA, the gene-targeting vectors for nab2 (PBANKA_1122000), tho4 (PBANKA_1230500), npl3 (PBANKA_0506600) and sr1 (PBANKA_1232100) were prepared by PCR. Briefly, the 5’ and 3’ flanking regions of the open reading frame (ORF) of target genes were amplified by PCR. The PCR products were annealed to either side of the human dihydrofolate reductase (hdhfr)-expressing cassette and amplified by PCR using gene-specific primers The gene-targeting vectors were introduced into the ORFs of target genes by double-crossover homologous recombination

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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