RMgmDB - Rodent Malaria genetically modified Parasites

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Summary

RMgm-1432

Malaria parasite	P. berghei
Genotype
Disrupted	Gene model (rodent): PBANKA_0417200; Gene model (P.falciparum): PF3D7_0904200; Gene product: PH domain-containing protein, putative (PH; PbPH; POM2; putative ookinete microneme protein, 2)
Phenotype	Gametocyte/Gamete; Fertilization and ookinete; Oocyst;

Last modified: 8 April 2016, 18:28

*RMgm-1432

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) : 27038925
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	Kou X; Cao Y
Name Group/Department	Department of Immunology
Name Institute	College of Basic Medical Sciences, China Medical University
City	Shenyang, Liaoning
Country	China
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Name of the mutant parasite
RMgm number	RMgm-1432
Principal name	Δpbph
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	Reduced (1.8 fold) production of gametocytes
Fertilization and ookinete	Reduced (34%) ookinete production in vitro
Oocyst	Reduced oocysts formation (the mean number of oocysts per midgut was reduced from 126.9 in those fed on wild-type parasites to 63.7 in those fed on Δpbph parasites).
Sporozoite	Not tested
Liver stage	Not tested
Additional remarks phenotype	Mutant/mutation Protein (function) The protein contains a signal peptide and a pleckstrin homology (PH) domain. PH domains are small modular domains that occur in a large variety of eukaryotic signaling proteins. The domains can bind to phosphatidylinositol in biological membranes and also to proteins such as protein kinase C. Through such interactions, PH domains recruit proteins to different membranes and target them to appropriate cellular compartments or enable them to interact with other components of a signal transduction pathway. Phenotype Reduced gametocyte, ookinete and oocyst production. See also RMgm-1292: An indepndent mutant lacking expression of PbPH (showing normal production of ookinetes/oocysts) Additional information Evidence is presented for expression in (male/female) gametocytes and ookinetes. IFA analysis provided evidence of surface location on gametes, zygotes and ookinetes. Other mutants RMgm-1292: An indepndent mutant lacking expression of PbPH (showing normal production of ookinetes/oocysts)

Disrupted: Mutant parasite with a disrupted gene

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Details of the target gene

Gene Model of Rodent Parasite

PBANKA_0417200

Gene Model P. falciparum ortholog

PF3D7_0904200

Gene product

PH domain-containing protein, putative

Gene product: Alternative name

PH; PbPH; POM2; putative ookinete microneme 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

eef1a

Selection (positive) procedure

pyrimethamine

Selection (negative) procedure

Additional remarks genetic modification

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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