| Synonymous Codon Usage Bias and Overexpression of a Synthetic Gene Encoding Interferon α2b in Yeast |
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| 作者姓名： | Bin FANG Bu-feng LIANG Guang-yuan HE |
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| 作者单位： | [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 |
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| 基金项目： | The National ‘973’ Basic Research Program (2002CB111302)，The National Natural Science Foundation of China (30370807) |
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| 摘 要： | 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.
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| 关 键 词： | Synthetic gene Interferon α2b Codon bias Pichia pastoris |
| 文章编号： | 1003-5125（2007）03-0226-07 |
| 修稿时间： | 2006-11-222007-01-17 |
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