RMgmDB - Rodent Malaria genetically modified Parasites

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Summary

RMgm-1435

Malaria parasite	P. berghei
Genotype
Tagged	Gene model (rodent): PBANKA_1222200; Gene model (P.falciparum): PF3D7_0808450; Gene product: protein MPODD, putative (mitochondrial protein ookinete developmental defect (MPODD)) Name tag: mCherry
Phenotype	Asexual bloodstage; Gametocyte/Gamete; Fertilization and ookinete; Sporozoite; Liver stage;

Last modified: 9 April 2016, 13:33

*RMgm-1435

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) : 27053680
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	Klug, D; Frischknecht F; Douglas RG
Name Group/Department	Integrative Parasitology, Center for Infectious Diseases
Name Institute	Heidelberg University Medical School
City	Heidelberg
Country	Germany
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Name of the mutant parasite
RMgm number	RMgm-1435
Principal name	mpodd::mCh
Alternative name
Standardized name
Is the mutant parasite cloned after genetic modification	Yes
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Phenotype
Asexual blood stage	The tagged protein was observed in all life stages with an apparent increased fluorescent signal in mosquito-stage parasites. MPODD::mCh localized to the mitochondrion at all life stages as seen by the distinct co-localization of the fusion protein with the dye MitoTracker.
Gametocyte/Gamete	The tagged protein was observed in all life stages with an apparent increased fluorescent signal in mosquito-stage parasites. MPODD::mCh localized to the mitochondrion at all life stages as seen by the distinct co-localization of the fusion protein with the dye MitoTracker.
Fertilization and ookinete	The tagged protein was observed in all life stages with an apparent increased fluorescent signal in mosquito-stage parasites. MPODD::mCh localized to the mitochondrion at all life stages as seen by the distinct co-localization of the fusion protein with the dye MitoTracker.
Oocyst	Not tested
Sporozoite	The tagged protein was observed in all life stages with an apparent increased fluorescent signal in mosquito-stage parasites. MPODD::mCh localized to the mitochondrion at all life stages as seen by the distinct co-localization of the fusion protein with the dye MitoTracker.
Liver stage	The tagged protein was observed in all life stages with an apparent increased fluorescent signal in mosquito-stage parasites. MPODD::mCh localized to the mitochondrion at all life stages as seen by the distinct co-localization of the fusion protein with the dye MitoTracker.
Additional remarks phenotype	Mutant/mutation The mutant expresses a C-terminal mCherry-tagged version of of MPODD. Protein (function) Bioinformatic analysis of MPODD revealed the presence of a TM domain as well as a mitochondrial targeting signal. Phenotype The tagged protein was observed in all life stages with an apparent increased fluorescent signal in mosquito-stage parasites. MPODD::mCh localized to the mitochondrion at all life stages as seen by the distinct co-localization of the fusion protein with the dye MitoTracker. Phenotype analyses of a mutant lacking expression of MPODD (RMgm-1434) showed the following: Normal blood stage development and gametocyte production. Aberrant ookinete development. Knockout parasites failed to develop mature, infectious ookinetes. mpodd(-) mutants typically resembled retort forms with the nucleus still positioned in the bulb-like extension of the zygote, suggesting an important role in final conversion from zygotes to ookinetes. No oocyst production. Additional information Other mutants A mutant lacking expression of MPODD (RMgm-1434)

Tagged: Mutant parasite with a tagged gene

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Details of the target gene

Gene Model of Rodent Parasite

PBANKA_1222200

Gene Model P. falciparum ortholog

PF3D7_0808450

Gene product

protein MPODD, putative

Gene product: Alternative name

mitochondrial protein ookinete developmental defect (MPODD)

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Details of the genetic modification

Name of the tag

mCherry

Details of tagging

C-terminal

Additional remarks: tagging

Commercial source of tag-antibodies

Type of plasmid/construct

(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

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

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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