RMgmDB - Rodent Malaria genetically modified Parasites

Summary

RMgm-1548

Malaria parasite	P. yoelii
Genotype
Tagged	Gene model (rodent): PY17X_1372600; Gene model (P.falciparum): Not available; Gene product: fam-b protein Name tag: EGFP
Phenotype	Asexual bloodstage;

Last modified: 15 August 2016, 15:16

*RMgm-1548

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) : 27503796
MR4 number
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Parent parasite used to introduce the genetic modification
Rodent Malaria Parasite	P. yoelii
Parent strain/line	P. y. yoelii 17X
Name parent line/clone	cl1.1
Other information parent line
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The mutant parasite was generated by
Name PI/Researcher	Siau, A; Preiser, P.R.
Name Group/Department	School of Biological Sciences
Name Institute	Nanyang Technological University
City	Singapore
Country	Singapore
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Name of the mutant parasite
RMgm number	RMgm-1548
Principal name	PY17X_1372600-GFP
Alternative name
Standardized name
Is the mutant parasite cloned after genetic modification	No
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Phenotype
Asexual blood stage	Exported protein: location in the cytoplasm of the infected red blood cell (punctate GFP pattern)
Gametocyte/Gamete	Not tested
Fertilization and ookinete	Not tested
Oocyst	Not tested
Sporozoite	Not tested
Liver stage	Not tested
Additional remarks phenotype	Mutant/mutation The mutant expresses a GFP-tagged version of the protein. The transgene is introduced on a plasmid and is expressed under the control of the constitutive eef1a promoter of P. berghei The protein was identified in a labelfree quantitative LC-MS/MS analysis of the whole proteome subcellular fractionation of erythrocytic P. yoelii at trophozoite and schizont stage with the aim to identify proteins involved in remodelling the host red blood cell. A total of 61 proteins were C-terminally tagged with GFP (expressed from episomes) followed by live-cell fluorescence microscopy analysis of the tagged protein in (uncloned) blood stages. Three different locations were distinguished: peripheral location (at the periphery of the parasite), location in the cytoplasm of the infected red blood cell (exported proteins) and an internal location Phenotype Exported protein: location in the cytoplasm of the infected red blood cell (punctate GFP pattern).

Tagged: Mutant parasite with a tagged gene

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Details of the target gene

Gene Model of Rodent Parasite

PY17X_1372600

Gene Model P. falciparum ortholog

Not available

Gene product

fam-b protein

Gene product: Alternative name

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Details of the genetic modification

Name of the tag

EGFP

Details of tagging

C-terminal

Additional remarks: tagging

Commercial source of tag-antibodies

Type of plasmid/construct

Circular plasmid

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

The mutant expresses a GFP-tagged version of the protein. The transgene is introduced on a plasmid and is expressed under the control of the constitutive eef1a promoter of P. berghei

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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