Abstract
Aprotinin is a small serine protease inhibitor used in human health. Spirodela were transformed, via Agrobacterium, with a synthetic gene encoding the mature aprotinin sequence and a signal peptide for secretion which was driven by the CaMV 35S promoter. A total of 25 transgenic Spirodela lines were generated and aprotinin production was confirmed by northern and western blot analyses. Expression levels of up to 3.7% of water soluble proteins were detected in the plant and 0.65 mg/l in the growth medium. In addition, immunoaffinity purification of the protein followed by amino acid sequencing confirmed the correct splicing of the aprotinin produced in Spirodela and secreted into the growth medium.
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Acknowledgements
This work was supported by Bayer BioScience. LemnaGene SA benefited from a Grant from ANVAR. We thank Isabelle Zanella-Cleon and Michel Becchi from the IFR 128 at the IBPC Lyon, France, for performing the amino acid sequence determination of the purified aprotinin.
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Rival, S., Wisniewski, JP., Langlais, A. et al. Spirodela (duckweed) as an alternative production system for pharmaceuticals: a case study, aprotinin. Transgenic Res 17, 503–513 (2008). https://doi.org/10.1007/s11248-007-9123-x
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DOI: https://doi.org/10.1007/s11248-007-9123-x