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A toolset of constitutive promoters for metabolic engineering of Rhodosporidium toruloides.
Nora, Luísa Czamanski; Wehrs, Maren; Kim, Joonhoon; Cheng, Jan-Fang; Tarver, Angela; Simmons, Blake A; Magnuson, Jon; Harmon-Smith, Miranda; Silva-Rocha, Rafael; Gladden, John M; Mukhopadhyay, Aindrila; Skerker, Jeffrey M; Kirby, James.
Afiliación
  • Nora LC; Ribeirão Preto Medical School, University of São Paulo, Av. Bandeirantes 3900, Ribeirão Prêto, São Paulo, CEP 14049-900, Brazil.
  • Wehrs M; DOE Joint BioEnergy Institute, 5885 Hollis St, Emeryville, CA, 94608, USA.
  • Kim J; Biological Systems and Engineering Division, Lawrence Berkeley National Laboratory, 1 Cyclotron Rd, Berkeley, CA, 94720, USA.
  • Cheng JF; DOE Joint BioEnergy Institute, 5885 Hollis St, Emeryville, CA, 94608, USA.
  • Tarver A; Biological Systems and Engineering Division, Lawrence Berkeley National Laboratory, 1 Cyclotron Rd, Berkeley, CA, 94720, USA.
  • Simmons BA; Institut für Genetik, Technische Universität Braunschweig, 38106, Brunswick, Germany.
  • Magnuson J; DOE Joint BioEnergy Institute, 5885 Hollis St, Emeryville, CA, 94608, USA.
  • Harmon-Smith M; Chemical and Biological Process Development Group, Pacific Northwest National Laboratory, 902 Battelle Blvd, Richland, WA, 99354, USA.
  • Silva-Rocha R; Department of Energy, Joint Genome Institute, 2800 Mitchell Drive, Walnut Creek, CA, 94598, USA.
  • Gladden JM; Environmental Genomics and Systems Biology Division, Lawrence Berkeley National Laboratory, 1 Cyclotron Rd, Berkeley, CA, 94720, USA.
  • Mukhopadhyay A; Department of Energy, Joint Genome Institute, 2800 Mitchell Drive, Walnut Creek, CA, 94598, USA.
  • Skerker JM; Environmental Genomics and Systems Biology Division, Lawrence Berkeley National Laboratory, 1 Cyclotron Rd, Berkeley, CA, 94720, USA.
  • Kirby J; DOE Joint BioEnergy Institute, 5885 Hollis St, Emeryville, CA, 94608, USA.
Microb Cell Fact ; 18(1): 117, 2019 Jun 29.
Article en En | MEDLINE | ID: mdl-31255171
BACKGROUND: Rhodosporidium toruloides is a promising host for the production of bioproducts from lignocellulosic biomass. A key prerequisite for efficient pathway engineering is the availability of robust genetic tools and resources. However, there is a lack of characterized promoters to drive expression of heterologous genes for strain engineering in R. toruloides. RESULTS: This data describes a set of native R. toruloides promoters, characterized over time in four different media commonly used for cultivation of this yeast. The promoter sequences were selected using transcriptional analysis and several of them were found to drive expression bidirectionally. Promoter expression strength was determined by measurement of EGFP and mRuby2 reporters by flow cytometry. A total of 20 constitutive promoters (12 monodirectional and 8 bidirectional) were found, and are expected to be of potential value for genetic engineering of R. toruloides. CONCLUSIONS: A set of robust and constitutive promoters to facilitate genetic engineering of R. toruloides is presented here, ranging from a promoter previously used for this purpose (P7, glyceraldehyde 3-phosphate dehydrogenase, GAPDH) to stronger monodirectional (e.g., P15, mitochondrial adenine nucleotide translocator, ANT) and bidirectional (e.g., P9 and P9R, histones H3 and H4, respectively) promoters. We also identified promoters that may be useful for specific applications such as late-stage expression (e.g., P3, voltage-dependent anion channel protein 2, VDAC2). This set of characterized promoters significantly expands the range of engineering tools available for this yeast and can be applied in future metabolic engineering studies.
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Rhodotorula / Regiones Promotoras Genéticas / Ingeniería Metabólica Idioma: En Revista: Microb Cell Fact Asunto de la revista: BIOTECNOLOGIA / MICROBIOLOGIA Año: 2019 Tipo del documento: Article País de afiliación: Brasil Pais de publicación: Reino Unido
Texto completo
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Rhodotorula / Regiones Promotoras Genéticas / Ingeniería Metabólica Idioma: En Revista: Microb Cell Fact Asunto de la revista: BIOTECNOLOGIA / MICROBIOLOGIA Año: 2019 Tipo del documento: Article País de afiliación: Brasil Pais de publicación: Reino Unido