Gene, 2000-03, Vol.245 (1), p.169-174
2000
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- Autor(en) / Beteiligte
- Titel
- Rapid, high-level expression of glycosylated rice α-amylase in transfected plants by an RNA viral vector
- Ist Teil von
- Gene, 2000-03, Vol.245 (1), p.169-174
- Ort / Verlag
- Netherlands: Elsevier B.V
- Erscheinungsjahr
- 2000
- Quelle
- Elsevier ScienceDirect Journals
- Beschreibungen/Notizen
- Tobamoviral vectors have been developed for the heterologous expression of glycoproteins in plants. The rice α-amylase gene (OS103) was placed under the transcriptional control of a tobamovirus subgenomic promoter in a RNA viral vector. One to two weeks after inoculation, transfected Nicotiana benthamiana plants accumulated glycosylated α-amylase to levels of at least 5% total soluble protein. The 46 kDa recombinant enzyme was purified, and its structural and biological properties were analyzed. Post-translational modifications of the secreted protein were compared to rice α-amylase expressed in amylolytic strains of Pichia pastoris and Saccharomyces cerevisiae. Endo-H analysis revealed that the α-amylase was moderately glycosylated in transfected plants and hyperglycosylated in yeast.
- Sprache
- Englisch
- Identifikatoren
- ISSN: 0378-1119eISSN: 1879-0038DOI: 10.1016/S0378-1119(00)00015-9Titel-ID: cdi_proquest_miscellaneous_70956353
- Format
- –
- Schlagworte
- alpha-Amylases - genetics, alpha-Amylases - metabolism, Amino Acid Sequence, Base Sequence, Blotting, Western, Gene Expression Regulation, Enzymologic, Genetic Vectors, Glycoprotein, Glycosylation, Molecular Sequence Data, Nicotiana benthamiana, Oryza - enzymology, Oryza - genetics, Oryza - virology, Oryza sativa, Pichia pastoris, Recombinant Fusion Proteins - genetics, Recombinant Fusion Proteins - metabolism, Saccharomyces cerevisiae, Subgenomic promoter, Tobamovirus, Tobamovirus - genetics, Tobamovirus - ultrastructure, Transfection