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Virologica Sinica, 22 (3) : 226-232, 2007
Research Article
Synonymous Codon Usage Bias and Overexpression of a Synthetic Gene Encoding Interferon α2b in Yeast
1. China-UK HUST-RRes Genetic Engineering and Genomics Joint Lab, Huazhong University of Science and Technology HUST, Wuhan 430074, China
2. Joint R&D Lab Suzhong Pharma Co.,Ltd, IHS, Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan 430071, China
To achieve higher level expression of Interferon α2b (IFN-α2b) in methylotrophic yeast (Pichia pastoris), a cDNA fragment coding for the mature IFN-α2b was designed and synthesized based on the synonymous codon bias of P. pastoris and optimized G+C content. The synthetic IFN-α2b was inserted into the secreted expression vector pPICZαA, and then integrated into P. pastoris GS115 genome by electroporation. Multi-copy integrants in the Mut+ recombinant P. pastoris strain were screened by high concentrations of Zeocin. 120 hours culturing allowed expression of the IFN-α2b transformant up to 810 mg/L as detected by SDS-PAGE and quantitative methods. In addition, Western blot analysis showed that the recombinant proteins had immunogenicity. The significant antiviral activity of the recombinant IFN-α2b protein was verified by WISH/ VSV system, which was 3.3×105 IU/mL.
  Published online: 20 Jun 2007
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