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High-level expression of active human alpha1-antitrypsin in transgenic tobacco chloroplasts

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Abstract

We have produced human alpha1-antitrypsin (A1AT), a major therapeutic protein, in genetically engineered tobacco plastids. Four different expression vectors have been evaluated which encode A1AT under the control of various 5′ and 3′ plastid expression elements. The use of heterologous promoter and terminator sequences derived from the corn and soybean plastid genomes leads to simpler and predictable recombinant genome patterns, avoiding unwanted recombination products between introduced and resident tobacco sequences. High level expression of unglycosylated A1AT, representing up to 2% of total soluble proteins, has been measured in leaves of transgenic tobacco lines. Some heterogeneity in the recombinant A1AT is detected after 2D protein separation, but the chloroplast-made protease inhibitors are fully active and bind to porcine pancreatic elastase.

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Authors and Affiliations

  1. Bayer BioScience, 14-20 Rue Pierre Baizet, BP 9163, 69263, Lyon cedex 09, France

    M. Nadai, J. Bally, M. Vitel, G. Tissot & M. Dubald

  2. Centre National de la Recherche Scientifique, Bayer CropScience Joint Laboratory, Unité Mixte de Recherche 5240, 69263, Lyon cedex 09, France

    C. Job

  3. Bayer BioScience, Technologiepark 38, 9052, Ghent, Belgium

    J. Botterman

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  1. M. Nadai
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  2. J. Bally
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  4. C. Job
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  6. J. Botterman
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Correspondence to M. Dubald.

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Nadai, M., Bally, J., Vitel, M. et al. High-level expression of active human alpha1-antitrypsin in transgenic tobacco chloroplasts. Transgenic Res 18, 173–183 (2009). https://doi.org/10.1007/s11248-008-9209-0

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