Improved transfection and new selectable markers for the rodent malaria parasite Plasmodium yoelii

Artemio M. Jongco; Li Min Ting; Vandana Thathy; Maria M. Mota; Kami Kim

doi:10.1016/j.molbiopara.2006.01.001

Improved transfection and new selectable markers for the rodent malaria parasite Plasmodium yoelii

Artemio M. Jongco, Li Min Ting, Vandana Thathy, Maria M. Mota, Kami Kim

Medicine

Research output: Contribution to journal › Article › peer-review

53 Scopus citations

Abstract

The rodent malaria Plasmodium yoelii is a useful model to study protective immunity to pre-erythrocytic stages of infection, pathogenesis of erythrocytic stages, and vaccine development. However, the utility of the P. yoelii model system has not been fully realized because transfection and genetic manipulation methodologies for this rodent species are less developed than that of another rodent species Plasmodium berghei. Here we report improved transfection efficiency using the AMAXA nucleofector® system compared to conventional transfection methodologies. We also show that heterologous promoters from P. berghei can be used to drive expression of a green fluorescent protein (GFP) reporter protein in P. yoelii. In an effort to develop additional selectable markers for this parasite, we also tested positive selectable markers that have been used successfully in P. falciparum and P. berghei. Human dihydrofolate reductase (hdhfr) and Toxoplasma gondii dihydrofolate reductase-thymidylate synthase (Tgdhfr-ts) conferred drug resistance to WR99210 and pyrimethamine, respectively, when introduced as episomes. These improvements should make genetic manipulation of P. yoelii more amenable and facilitate further studies of host-parasite interactions using this attractive rodent model.

Original language	English (US)
Pages (from-to)	242-250
Number of pages	9
Journal	Molecular and Biochemical Parasitology
Volume	146
Issue number	2
DOIs	https://doi.org/10.1016/j.molbiopara.2006.01.001
State	Published - Apr 2006

Keywords

FACS
GFP
Nucleofector®
Plasmodium yoelii
Transfection

ASJC Scopus subject areas

Parasitology
Molecular Biology

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

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10.1016/j.molbiopara.2006.01.001

Cite this

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title = "Improved transfection and new selectable markers for the rodent malaria parasite Plasmodium yoelii",

abstract = "The rodent malaria Plasmodium yoelii is a useful model to study protective immunity to pre-erythrocytic stages of infection, pathogenesis of erythrocytic stages, and vaccine development. However, the utility of the P. yoelii model system has not been fully realized because transfection and genetic manipulation methodologies for this rodent species are less developed than that of another rodent species Plasmodium berghei. Here we report improved transfection efficiency using the AMAXA nucleofector{\textregistered} system compared to conventional transfection methodologies. We also show that heterologous promoters from P. berghei can be used to drive expression of a green fluorescent protein (GFP) reporter protein in P. yoelii. In an effort to develop additional selectable markers for this parasite, we also tested positive selectable markers that have been used successfully in P. falciparum and P. berghei. Human dihydrofolate reductase (hdhfr) and Toxoplasma gondii dihydrofolate reductase-thymidylate synthase (Tgdhfr-ts) conferred drug resistance to WR99210 and pyrimethamine, respectively, when introduced as episomes. These improvements should make genetic manipulation of P. yoelii more amenable and facilitate further studies of host-parasite interactions using this attractive rodent model.",

keywords = "FACS, GFP, Nucleofector{\textregistered}, Plasmodium yoelii, Transfection",

author = "Jongco, {Artemio M.} and Ting, {Li Min} and Vandana Thathy and Mota, {Maria M.} and Kami Kim",

note = "Funding Information: AMJ was partially supported by NIH Medical Scientist Training Program grant GM07288. VT was partially supported by NIH Training grant T32A107506 awarded to the Albert Einstein College of Medicine. This work was supported by a Burroughs Wellcome Fund New Initiatives in Malaria Research Award, US Army Research grant PR043176 and a Marion Bessin Liver Research Center Pilot and Feasibility grant awarded to KK. We thank Chris Janse and Andy Waters for providing plasmids, helpful advice, and communication of unpublished results prior to publication.",

year = "2006",

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doi = "10.1016/j.molbiopara.2006.01.001",

language = "English (US)",

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AU - Jongco, Artemio M.

AU - Ting, Li Min

AU - Thathy, Vandana

AU - Mota, Maria M.

AU - Kim, Kami

N1 - Funding Information: AMJ was partially supported by NIH Medical Scientist Training Program grant GM07288. VT was partially supported by NIH Training grant T32A107506 awarded to the Albert Einstein College of Medicine. This work was supported by a Burroughs Wellcome Fund New Initiatives in Malaria Research Award, US Army Research grant PR043176 and a Marion Bessin Liver Research Center Pilot and Feasibility grant awarded to KK. We thank Chris Janse and Andy Waters for providing plasmids, helpful advice, and communication of unpublished results prior to publication.

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N2 - The rodent malaria Plasmodium yoelii is a useful model to study protective immunity to pre-erythrocytic stages of infection, pathogenesis of erythrocytic stages, and vaccine development. However, the utility of the P. yoelii model system has not been fully realized because transfection and genetic manipulation methodologies for this rodent species are less developed than that of another rodent species Plasmodium berghei. Here we report improved transfection efficiency using the AMAXA nucleofector® system compared to conventional transfection methodologies. We also show that heterologous promoters from P. berghei can be used to drive expression of a green fluorescent protein (GFP) reporter protein in P. yoelii. In an effort to develop additional selectable markers for this parasite, we also tested positive selectable markers that have been used successfully in P. falciparum and P. berghei. Human dihydrofolate reductase (hdhfr) and Toxoplasma gondii dihydrofolate reductase-thymidylate synthase (Tgdhfr-ts) conferred drug resistance to WR99210 and pyrimethamine, respectively, when introduced as episomes. These improvements should make genetic manipulation of P. yoelii more amenable and facilitate further studies of host-parasite interactions using this attractive rodent model.

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Improved transfection and new selectable markers for the rodent malaria parasite Plasmodium yoelii

Abstract

Keywords

ASJC Scopus subject areas

UN SDGs

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