Wolffia arrhiza as a promising producer of recombinant hirudin
| Autor: | A. P. Firsov, Lyubov Shaloiko, Pavel Khvatkov, Mariya Chernobrovkina, Sergey Dolgov, Alexander Pushin, Anastasiya Shvedova, Oleg Kozlov |
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| Jazyk: | angličtina |
| Rok vydání: | 2021 |
| Předmět: |
biology
Transgene Genetically modified crops Environmental Science (miscellaneous) biology.organism_classification Agricultural and Biological Sciences (miscellaneous) Wolffia arrhiza Wolffia law.invention Transformation (genetics) Biochemistry law Recombinant DNA Original Article Target protein Biotechnology Transformation efficiency |
| Zdroj: | 3 Biotech |
| Popis: | The production of recombinant proteins in transgenic plants is becoming an increasingly serious alternative to classical biopharming methods as knowledge about this process grows. Wolffia arrhiza, an aquatic plant unique in its anatomy, is a promising expression system that can grow in submerged culture in bioreactors. In our study 8550 explants were subjected to Agrobacterium-mediated transformation, and 41 independent hygromycin-resistant Wolffia lines were obtained, with the transformation efficiency of 0.48%. 40 of them contained the hirudin-1 gene (codon-optimized for expression in plants) and were independent lines of nuclear-transformed Wolffia, the transgenic insertion has been confirmed by PCR and Southern blot analysis. We have analyzed the accumulation of the target protein and its expression has been proven in three transgenic lines. The maximum accumulation of recombinant hirudin was 0.02% of the total soluble protein, which corresponds to 775.5 ± 111.9 ng g(−1) of fresh weight of the plant. The results will be used in research on the development of an expression system based on Wolffia plants for the production of hirudin and other recombinant pharmaceutical proteins. |
| Databáze: | OpenAIRE |
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