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| Secretory Expression and Purification of HSA-NIFN Fusion Protein in Pichia pastoris |
| TIAN Shuo1,YAO Wen-bin1,ZHOU Min-yi2,LI Jie2,SONG Wen-jin2,XU Chen2 |
1.School of Life Science & Technology，China Pharmaceutical University，Nanjing 210009，China
2.Beijing Tri-prime Genetic Engineering Co．Ltd．，Beijing 102600，China |
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Abstract Abstract HSA-NIFN gene was got by total gene synthesis technology in vitro. The NIFN-HSA gene was inserted into Pichia pastoris express vector pPIC3.5. The recombinant plasmid pPIC3.5/HSA-NIFN was linearized by restriction enzyme Sal I and then transformed into Pichia pastoris GS115 by electroporation. The recombinant strains confirmed by PCR analysis and were induced by methanol to express fusion protein HSA-NIFN. SDS-PAGE , Western blot and MALDI-TOF-MS analysis of the fusion protein showed that the expressed fusion protein HSA-NIFN with 86369Da molecular weight had the antigenicity of HSA.The putity of HSA-NIFN was higher than 90% after Blue Sepharose FF and CM Sepharose FF, and the biological activity determined by WISH-VSV system was more than 7.75±0.39×106 IU/mg.
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Received: 12 May 2010
Published: 12 June 2010
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