Lozano, Jean-Claude, Schatt, Philippe, Botebol, Hugo, Verge, Valerie, Lesuisse, Emmanuel, Blain, Stephane, Carré, Isabelle A. and Bouget, Francois-Yves (2014) Efficient gene targeting and foreign DNA removal by homologous recombination in the picoeukaryote Ostreococcus. Plant Journal, Volume 78 (Number 6). pp. 1073-1083. ISSN 0960-7412.

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Abstract

With fewer than 8000 genes and a minimalist cellular organization, the green picoalga Ostreococcus tauri is one of the simplest photosynthetic eukaryotes. Ostreococcus tauri contains many plant-specific genes but exhibits a very low gene redundancy. The haploid genome is extremely dense with few repeated sequences and rare transposons. Thanks to the implementation of genetic transformation and vectors for inducible overexpression/knockdown this picoeukaryotic alga has emerged in recent years as a model organism for functional genomics analyses and systems biology. Here we report the development of an efficient gene targeting technique which we use to knock out the nitrate reductase and ferritin genes and to knock in a luciferase reporter in frame to the ferritin native protein. Furthermore, we show that the frequency of insertion by homologous recombination is greatly enhanced when the transgene is designed to replace an existing genomic insertion. We propose that a natural mechanism based on homologous recombination may operate to remove inserted DNA sequences from the genome.

Item Type:	Journal Article
Subjects:	Q Science > Q Science (General) Q Science > QK Botany Q Science > QR Microbiology > QR355 Virology
Divisions:	Faculty of Science, Engineering and Medicine > Science > Life Sciences (2010- )
Library of Congress Subject Headings (LCSH):	Green algae, Plant cytogenetics, Genetic recombination, Gene targeting, Microalgae, Genetic transformation
Journal or Publication Title:	Plant Journal
Publisher:	Wiley-Blackwell Publishing Ltd.
ISSN:	0960-7412
Official Date:	June 2014
Dates:	Date Event June 2014 Published 11 June 2014 Available 31 March 2014 Accepted 18 July 2013 Submitted
Volume:	Volume 78
Number:	Number 6
Page Range:	pp. 1073-1083
Status:	Peer Reviewed
Publication Status:	Published
Access rights to Published version:	Restricted or Subscription Access
Date of first compliant deposit:	28 July 2016
Date of first compliant Open Access:	28 July 2016

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