2A and the auxin-based degron system facilitate control of protein levels in Plasmodium falciparum.
Analysis of gene function in Plasmodium falciparum, the most important human malaria parasite, is restricted by the lack of robust and simple reverse genetic tools. Approaches to manipulate protein levels post-translationally are powerful tools to study protein-off effects especially in the haploid...
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2013
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oai:doaj.org-article:3b007483133b4e5281463b779559f6082021-11-18T08:46:56Z2A and the auxin-based degron system facilitate control of protein levels in Plasmodium falciparum.1932-620310.1371/journal.pone.0078661https://doaj.org/article/3b007483133b4e5281463b779559f6082013-01-01T00:00:00Zhttps://www.ncbi.nlm.nih.gov/pmc/articles/pmid/24236031/pdf/?tool=EBIhttps://doaj.org/toc/1932-6203Analysis of gene function in Plasmodium falciparum, the most important human malaria parasite, is restricted by the lack of robust and simple reverse genetic tools. Approaches to manipulate protein levels post-translationally are powerful tools to study protein-off effects especially in the haploid malaria parasite where genetic knockouts of essential genes are lethal. We investigated if the auxin-inducible degron system is functional in P. falciparum and found that degron-tagged yellow fluorescent protein levels were efficiently reduced upon addition of auxin which otherwise had no effect on parasite viability. The genetic components required in this conditional approach were co-expressed in P. falciparum by applying the small peptide 2A. 2A is a self-processing peptide from Foot-And-Mouth Disease virus that allows the whole conditional system to be accommodated on a single plasmid vector and ensures stoichiometric expression levels.Andrea KreidenweissAnnika V HopkinsBenjamin MordmüllerPublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 8, Iss 11, p e78661 (2013) |
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Medicine R Science Q Andrea Kreidenweiss Annika V Hopkins Benjamin Mordmüller 2A and the auxin-based degron system facilitate control of protein levels in Plasmodium falciparum. |
description |
Analysis of gene function in Plasmodium falciparum, the most important human malaria parasite, is restricted by the lack of robust and simple reverse genetic tools. Approaches to manipulate protein levels post-translationally are powerful tools to study protein-off effects especially in the haploid malaria parasite where genetic knockouts of essential genes are lethal. We investigated if the auxin-inducible degron system is functional in P. falciparum and found that degron-tagged yellow fluorescent protein levels were efficiently reduced upon addition of auxin which otherwise had no effect on parasite viability. The genetic components required in this conditional approach were co-expressed in P. falciparum by applying the small peptide 2A. 2A is a self-processing peptide from Foot-And-Mouth Disease virus that allows the whole conditional system to be accommodated on a single plasmid vector and ensures stoichiometric expression levels. |
format |
article |
author |
Andrea Kreidenweiss Annika V Hopkins Benjamin Mordmüller |
author_facet |
Andrea Kreidenweiss Annika V Hopkins Benjamin Mordmüller |
author_sort |
Andrea Kreidenweiss |
title |
2A and the auxin-based degron system facilitate control of protein levels in Plasmodium falciparum. |
title_short |
2A and the auxin-based degron system facilitate control of protein levels in Plasmodium falciparum. |
title_full |
2A and the auxin-based degron system facilitate control of protein levels in Plasmodium falciparum. |
title_fullStr |
2A and the auxin-based degron system facilitate control of protein levels in Plasmodium falciparum. |
title_full_unstemmed |
2A and the auxin-based degron system facilitate control of protein levels in Plasmodium falciparum. |
title_sort |
2a and the auxin-based degron system facilitate control of protein levels in plasmodium falciparum. |
publisher |
Public Library of Science (PLoS) |
publishDate |
2013 |
url |
https://doaj.org/article/3b007483133b4e5281463b779559f608 |
work_keys_str_mv |
AT andreakreidenweiss 2aandtheauxinbaseddegronsystemfacilitatecontrolofproteinlevelsinplasmodiumfalciparum AT annikavhopkins 2aandtheauxinbaseddegronsystemfacilitatecontrolofproteinlevelsinplasmodiumfalciparum AT benjaminmordmuller 2aandtheauxinbaseddegronsystemfacilitatecontrolofproteinlevelsinplasmodiumfalciparum |
_version_ |
1718421315773267968 |