Expression and recovery of biologically active recombinant Apolipoprotein AIMilano from transgenic safflower (Carthamus tinctorius) seeds
| Autor: | Maurice M. Moloney, Jean-Claude Tardif, Elizabeth Wanda Murray, Reid Alexandra, David Busseuil, Cory Nykiforuk, Joseph Boothe, Yin Shen, Eric Rhéaume |
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| Rok vydání: | 2011 |
| Předmět: |
biology
Apolipoprotein B Cholesterol Carthamus food and beverages Plant Science biology.organism_classification Fusion protein In vitro law.invention chemistry.chemical_compound chemistry Biochemistry law In vivo Phosphatidylcholine Recombinant DNA biology.protein Agronomy and Crop Science Biotechnology |
| Zdroj: | Plant Biotechnology Journal. 9:250-263 |
| ISSN: | 1467-7644 |
| DOI: | 10.1111/j.1467-7652.2010.00546.x |
| Popis: | Apolipoprotein AI Milano (ApoAI(Milano) ) was expressed as a fusion protein in transgenic safflower seeds. High levels of expression corresponding to 7 g of ApoAI(Milano) per kilogram of seed have been identified in a line selected for commercialization. The ApoAI(Milano) fusion protein was extracted from seed using an oilbody-based process and matured in vitro prior to final purification. This yielded a Des-1,2-ApoAI(Milano) product which was confirmed by biochemical characterization including immunoreactivity against ApoAI antibodies, isoelectric point, N-terminal sequencing and electrospray mass spectrometry. Purified Des-1,2-ApoAI(Milano) readily associated with dimyristoylphosphatidylcholine in clearance assays comparable to Human ApoAI. Its biological activity was assessed by cholesterol efflux assays using Des-1,2-ApoAI(Milano) :1-palmitoyl-2-oleoyl phosphatidylcholine complexes in vitro and in vivo. This study has established that high levels of biologically functional ApoAI(Milano) can be produced using a plant-based expression system. |
| Databáze: | OpenAIRE |
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