Simplified transformation of ostreococcus tauri using polyethylene glycol

Sanchez, F; Geffroy, S; Norest, M; Yau, S; Moreau, H; Grimsley, N

Simplified transformation of ostreococcus tauri using polyethylene glycol

Sanchez, F Geffroy, S

Norest, M Yau, S Moreau, H Grimsley, N

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Publication Type:: Journal Article
Citation:: Genes, 2019, 10 (5)
Issue Date:: 2019-05-01

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Field	Value	Language
dc.contributor.author	Sanchez, F	en_US
dc.contributor.author	Geffroy, S https://orcid.org/0000-0001-6247-5536	en_US
dc.contributor.author	Norest, M	en_US
dc.contributor.author	Yau, S	en_US
dc.contributor.author	Moreau, H	en_US
dc.contributor.author	Grimsley, N	en_US
dc.date.available	2019-05-21	en_US
dc.date.issued	2019-05-01	en_US
dc.identifier.citation	Genes, 2019, 10 (5)	en_US
dc.identifier.uri	http://hdl.handle.net/10453/137992
dc.description.abstract	© 2019 by the authors. Licensee MDPI, Basel, Switzerland. Ostreococcustauri is an easily cultured representative of unicellular algae (class Mamiellophyceae) that abound in oceans worldwide. Eight complete 13–22 Mb genomes of phylogenetically divergent species within this class are available, and their DNA sequences are nearly always present in metagenomic data produced from marine samples. Here we describe a simplified and robust transformation protocol for the smallest of these algae (O. tauri). Polyethylene glycol (PEG) treatment was much more effcient than the previously described electroporation protocol. Short (2 min or less) incubation times in PEG gave >104 transformants per microgram DNA. The time of cell recovery after transformation could be reduced to a few hours, permitting the experiment to be done in a day rather than overnight as used in previous protocols. DNA was randomly inserted in the O. tauri genome. In our hands PEG was 20–40-fold more effcient than electroporation for the transformation of O. tauri, and this improvement will facilitate mutagenesis of all of the dispensable genes present in the tiny O. tauri genome.	en_US
dc.relation.ispartof	Genes	en_US
dc.relation.isbasedon	10.3390/genes10050399	en_US
dc.subject.mesh	Polyethylene Glycols	en_US
dc.subject.mesh	Phylogeny	en_US
dc.subject.mesh	Transformation, Genetic	en_US
dc.subject.mesh	Base Sequence	en_US
dc.subject.mesh	Genome	en_US
dc.subject.mesh	Genetic Variation	en_US
dc.subject.mesh	Chlorophyta	en_US
dc.title	Simplified transformation of ostreococcus tauri using polyethylene glycol	en_US
dc.type	Journal Article
utslib.citation.volume	5	en_US
utslib.citation.volume	10	en_US
utslib.for	0604 Genetics	en_US
pubs.embargo.period	Not known	en_US
pubs.organisational-group	/University of Technology Sydney
pubs.organisational-group	/University of Technology Sydney/Faculty of Science
utslib.copyright.status	open_access
pubs.issue	5	en_US
pubs.publication-status	Published	en_US
pubs.volume	10	en_US

Abstract:

© 2019 by the authors. Licensee MDPI, Basel, Switzerland. Ostreococcustauri is an easily cultured representative of unicellular algae (class Mamiellophyceae) that abound in oceans worldwide. Eight complete 13–22 Mb genomes of phylogenetically divergent species within this class are available, and their DNA sequences are nearly always present in metagenomic data produced from marine samples. Here we describe a simplified and robust transformation protocol for the smallest of these algae (O. tauri). Polyethylene glycol (PEG) treatment was much more effcient than the previously described electroporation protocol. Short (2 min or less) incubation times in PEG gave >104 transformants per microgram DNA. The time of cell recovery after transformation could be reduced to a few hours, permitting the experiment to be done in a day rather than overnight as used in previous protocols. DNA was randomly inserted in the O. tauri genome. In our hands PEG was 20–40-fold more effcient than electroporation for the transformation of O. tauri, and this improvement will facilitate mutagenesis of all of the dispensable genes present in the tiny O. tauri genome.

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http://hdl.handle.net/10453/137992