REVIEW ARTICLE


High Stability of Recombinant Proteins Expressed in Tobacco Chloroplasts



Andrea Molina, Jon Veramendi*
Instituto de Agrobiotecnología, Universidad Pública de Navarra-CSIC-Gobierno de Navarra, Campus de Arrosadía, 31006 Pamplona, Spain


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Creative Commons License
© 2009 Molina and Veramendi

open-access license: This is an open access article distributed under the terms of the Creative Commons Attribution 4.0 International Public License (CC-BY 4.0), a copy of which is available at: (https://creativecommons.org/licenses/by/4.0/legalcode). This license permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

* Address correspondence to this author at the Instituto de Agrobiotecnología, Universidad Pública de Navarra-CSIC-Gobierno de Navarra, Campus de Arrosadía, 31006 Pamplona, Spain; Tel: +34-948-168033; Fax: +34-948-232191; E-mail: jon@unavarra.es


Abstract

The green fluorescent protein and the non-toxic subunit of cholera toxin were expressed in tobacco chloroplasts. Both recombinant proteins accumulated to levels greater than 20% of the total soluble protein. We did not observe light-dependent induction of gene expression despite employing the promoter and 5´-UTR from the psbA gene. Both proteins were stable in young and old leaves. The estimated half-life, determined by pulse-chase labeling, was greater than 48 h. Freeze-drying of pulverized leaves supplied an easy method of post-harvest handling with no significant loss of the recombinant protein after 7 months of storage at room temperature. These data suggest that both proteins are good candidates for the expression of fusion proteins (e.g. with antigens) in tobacco chloroplasts.

Keywords: Chloroplast transformation, protein stability, CTB, GFP.