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Plant Secondary Metabolism Engineering pp 275–290Cite as

Fusion with Fluorescent Proteins for Subcellular Localization of Enzymes Involved in Plant Alkaloid Biosynthesis

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Part of the book series: Methods in Molecular Biology ((MIMB,volume 643))

Abstract

To establish the role in alkaloid metabolism of candidate genes identified in silico or by Omics approaches, it may be essential to determine the subcellular localization of the encoded proteins. The fusion with fluorescent proteins (FP) may now be used as a quite effective and reliable tool to investigate this question. The methodology involves the choice of the FP, the design and production of the appropriate FP fusions, and the use of a transient or stable transformation protocol applied to a homologous or heterologous plant system. This chapter describes the application of this methodology to an enzyme involved in indole alkaloid biosynthesis, with general considerations on the development of the approach.

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Acknowledgements

This work was supported by: (i) Programa Operacional Ciência e Inovação (POCI 2010) financed by European Union and Fundação para a Ciência e Tecnologia (FCT) from the Portuguese Government; (ii) the FCT grants POCTI/BIO/38369/2001 and SFRH/BPD/20669/2004; (iii) Scientific Mecenate award from Grupo Jerónimo Martins SA.

Author information

Authors and Affiliations

  1. IBMC – Institute of Cellular and Molecular Biology, Porto University, Porto, Portugal

    Patrícia Duarte

  2. Institute of Biology, University of Leiden, Leiden, The Netherlands

    Johan Memelink

  3. IBMC – Institute of Molecular and Cellular Biology and Department of Biology of the Faculty of Sciences, University of Porto, Porto, Portugal

    Mariana Sottomayor

Authors
  1. Patrícia Duarte
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  2. Mariana Sottomayor
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Editors and Affiliations

  1. Rio Grande do Sul, Centro de Biotecnologia, Universidade Federal do, Porto Alegre, Rio Grande do Sul, Brazil

    Arthur Germano Fett-Neto

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Duarte, P., Memelink, J., Sottomayor, M. (2010). Fusion with Fluorescent Proteins for Subcellular Localization of Enzymes Involved in Plant Alkaloid Biosynthesis. In: Fett-Neto, A. (eds) Plant Secondary Metabolism Engineering. Methods in Molecular Biology, vol 643. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-60761-723-5_19

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  • DOI: https://doi.org/10.1007/978-1-60761-723-5_19

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  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-60761-722-8

  • Online ISBN: 978-1-60761-723-5

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