Production of native protein by using Synechocystis sp. PCC6803 DnaB mini-intein in Escherichia coli
| Autor: | Juan Yan, Gang Shi, Shou-Chun Chen, Xu Qi, Yu-Wang Tian, Shou-Sheng Yan |
|---|---|
| Rok vydání: | 2005 |
| Předmět: |
Genetic Vectors
Protein tag medicine.disease_cause Antiviral Agents Vesicular stomatitis Indiana virus Inteins FLAG-tag Protein purification Protein A/G Escherichia coli medicine Humans Cloning Molecular Expression vector Base Sequence biology Synechocystis Interferon-alpha Molecular biology Recombinant Proteins Interferon Type I biology.protein Protein G Intein Biotechnology |
| Zdroj: | Protein Expression and Purification. 40:340-345 |
| ISSN: | 1046-5928 |
| DOI: | 10.1016/j.pep.2004.12.021 |
| Popis: | To directly express native recombinant proteins in Escherichia coli, a new expression vector pSB was constructed using Ssp DnaB mini-intein. Using the vector, native proteins could be produced with the help of C-terminal self-cleavage of the intein. In this study, we cloned hIFNalpha-4 gene into pSB and used E. coli strain Origami B (DE3) as the host. Expression experiments were carried out both in Shake flasks and a 5 L bioreactor. The results indicated hIFNalpha-4 could be expressed in the form of soluble protein with correct folding in E. coli. The maximal hIFNalpha-4 content was 21.7% of total protein, and the antiviral activity of the protein was 1.2x10(8 )IU mg(-1). Overall, good effects were achieved with this system. This intein-mediated protein expression system opens up a useful method for production of native recombinant protein in E. coli. |
| Databáze: | OpenAIRE |
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