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Recombinant Gene Expression pp 209–223Cite as

Gene Transfer and Expression of Recombinant Proteins in Moderately Halophilic Bacteria

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Part of the Methods in Molecular Biology book series (MIMB,volume 267)

Abstract

Moderately halophilic bacteria (MHB) of the genera Halomonas and Chromohalobacter have been used as hosts for the expression of heterologous proteins of biotechnological interest, thus expanding their potential to be used as cell factories for various applications. This chapter deals with the methodology for the construction of recombinant plasmids, their transfer to a number of MHB, and the assaying of the corresponding heterologous proteins activity. The transferred genes include (1) inaZ, encoding the ice nucleation protein of the plant pathogen Pseudomonas syringae, (2) gfp, encoding a green fluorescent protein from the marine bioluminescent jellyfish Aequorea victoria, and (3) the α -amylase gene from the hyperthermophilic archeon Pyrococcus woesei. Vector pHS15, which was designed for expression of heterologous proteins in both E. coli and MHB, was used for the subcloning and transfer of the above genes. The recombinant constructs were introduced to MHB by assisted conjugal transfer from E. coli donors. The expression and function of the recombinant proteins in the MHB transconjugants is described.

Key Words

  • Halophilic bacteria
  • Halomonadaceae
  • Chromohalobacter
  • GFP
  • α-amylase
  • inaZ

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Author information

Authors and Affiliations

  1. Department of Chemistry, University of Ioannina, Ioannina, Greece

    Amalia S. Afendra & Constantin Drainas

  2. Department of Microbiology and Parasitology, Faculty of Pharmacy, University of Seville, Sevilla, España

    Carmen Vargas & Joaquín J. Nieto

Authors
  1. Amalia S. Afendra
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  2. Carmen Vargas
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  3. Joaquín J. Nieto
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  4. Constantin Drainas
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Editor information

Editors and Affiliations

  1. Centro de Investigación en Biotecnología, UAEM, Cuernavaca, Morelos, México

    Paulina Balbás

  2. Virginia Polytechnic Institute and State University, Blacksburg, VA

    Argelia Lorence

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© 2004 Humana Press Inc., Totowa, NJ

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Afendra, A.S., Vargas, C., Nieto, J.J., Drainas, C. (2004). Gene Transfer and Expression of Recombinant Proteins in Moderately Halophilic Bacteria. In: Balbás, P., Lorence, A. (eds) Recombinant Gene Expression. Methods in Molecular Biology, vol 267. Humana Press. https://doi.org/10.1385/1-59259-774-2:209

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  • DOI: https://doi.org/10.1385/1-59259-774-2:209

  • Publisher Name: Humana Press

  • Print ISBN: 978-1-58829-262-9

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