Yan Shou-Sheng, Yan Juan, Shi Gang, Xu Qi, Chen Shou-Chun, Tian Yu-Wang
School of Pharmaceutical Hunan College of TCM, Changsha, Hunan, China.
Protein Expr Purif. 2005 Apr;40(2):340-5. doi: 10.1016/j.pep.2004.12.021.
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.
