Recombinant Baculovirus Vectors Expressing Glutathione–S–Transferase Fusion Proteins
| Autor: | Ian M. Jones, Jeremy B. M. Jowett, Anthony H. Davies |
|---|---|
| Rok vydání: | 1993 |
| Předmět: |
Baculoviridae
Recombinant Fusion Proteins Genetic Vectors Molecular Sequence Data Retroviridae Proteins Biomedical Engineering Gene Expression Heterologous Bioengineering Moths Biology Recombinant virus Applied Microbiology and Biotechnology Cell Line law.invention Fusion gene law Animals Amino Acid Sequence Glutathione Transferase Membrane Glycoproteins Base Sequence Structural gene biology.organism_classification Fusion protein Molecular biology Biochemistry Cell culture Recombinant DNA Molecular Medicine Biotechnology |
| Zdroj: | Nature Biotechnology. 11:933-936 |
| ISSN: | 1546-1696 1087-0156 |
| Popis: | Recombinant baculoviruses are a popular means of producing heterologous protein in eukaryotic cells. Purification of recombinant proteins away from the insect cell background can, however, remain an obstacle for many developments. Recently, prokaryotic fusion protein expression systems have been developed allowing single-step purification of the heterologous protein and specific proteolytic cleavage of the affinity tag moiety from the desired antigen. Here we report the introduction of these attributes to the baculovirus system. "Baculo-GEX" vectors enable baculovirus production of fusion proteins with the above advantages, but in a eukaryotic post-translational processing environment. Glutathione-S-transferase (GST) fusions are stable cytoplasmic proteins in insect cells and may therefore be released by sonication alone, avoiding the solubility problems and detergent requirements of bacterial systems. Thus large amounts of authentic antigen may be purified in a single, non-denaturing step. |
| Databáze: | OpenAIRE |
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