title: Nuclear transformation and functional gene expression in the oleaginous microalga Monoraphidium neglectum.
abstract: Photosynthetic microalgae hold great promise as non-food feedstocks for the sustainable production of a range of bio-products and genetic engineering is an increasingly important strategy to improve the natural cellular features. For the vast majority of microalgal strains, however, genetic engineering is not yet possible and the establishment of efficient genetic tools for a broad range of microalgal species is an important task to reach biotechnological success. Stable DNA transformation is one of the crucial steps for most genetic engineering approaches. In this context, we report the first successful and stable nuclear genetic transformation of the biotechnologically promising oleaginous microalga M. neglectum. Transformation was achieved by electroporation and the efficiency could progressively be improved by implementation of an additional cell pretreatment step, aiming at weakening the rigid natural cell wall. Furthermore, a first reporter transgene was established for M. neglectum. As a result of this work, genetic engineering strategies now become accessible which are required to successfully establish M. neglectum as a novel host for biotechnological applications.
authors.affiliation: Algae Biotechnology and Bioenergy, Faculty of Biology, Center for Biotechnology (CeBiTec), Bielefeld University, 33615 Bielefeld, Germany.
authors.forename: Daniel
authors.initials: D
authors.lastname: Jaeger
authors.affiliation: Biomolecular Photonics, Faculty of Physics, Bielefeld University, 33615 Bielefeld, Germany.
authors.forename: Wolfgang
authors.initials: W
authors.lastname: Hübner
authors.affiliation: Biomolecular Photonics, Faculty of Physics, Bielefeld University, 33615 Bielefeld, Germany.
authors.forename: Thomas
authors.initials: T
authors.lastname: Huser
authors.affiliation: Algae Biotechnology and Bioenergy, Faculty of Biology, Center for Biotechnology (CeBiTec), Bielefeld University, 33615 Bielefeld, Germany.
authors.forename: Jan H
authors.initials: JH
authors.lastname: Mussgnug
authors.affiliation: Algae Biotechnology and Bioenergy, Faculty of Biology, Center for Biotechnology (CeBiTec), Bielefeld University, 33615 Bielefeld, Germany. Electronic address: olaf.kruse@uni-bielefeld.de.
authors.forename: Olaf
authors.initials: O
authors.lastname: Kruse
mesh_terms: D001709:Biotechnology; D000460:Chlorophyta / Q000235:genetics* / Q000378:metabolism; D018274:Electroporation / Q000379:methods; D008164:Luminescent Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D058086:Microalgae / Q000235:genetics* / Q000378:metabolism; D010957:Plasmids / Q000235:genetics; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D014170:Transformation, Genetic / Q000235:genetics*
chemical_list: D008164:Luminescent Proteins; D011994:Recombinant Proteins
keywords: Electroporation; Fluorescent proteins; Microalgae; Monoraphidium neglectum; Nuclear transformation