The fused gene (IFN-TFN) of TFN (transferring N-terminal half-molecule) gene and IFN (interferon) gene was amplified by multiple PCR.The fused gene and TFN gene was inserted into pPIC9 vector. The recombinant plasmid pPIC9-IFN-TFN and pPIC9-TFN were transformed into Pichia pastoris GS115 by PEG. After being induced by methanol, the target proteins were expressed in ferment supernatant at high level. The recombinant fused protein IFN-TFN and recombinant TFN with purity respectively being higher than 93% and 95% were finally obtained after purification through two-step chromatography: SP Sepharose Fast Flow and Phenyl Sepharose Fast Flow. According to in vitro bioactivity assay, the fused protein IFN-TFN had antiviral activity but which was much lower than the natural IFN. Fe3+ saturation study confirmed that the recombinant IFN-TFN was able to bind Fe3+ as the recombinant TFN did. It was shown that TFN could be used as the transcellar carrier of IFN.
LI Xiao-Jing, ZHANG Hao, XUE Chong, LI Yan-Ying, CHEN Jing, MIAO Lin, FANG Hong-Qing, CHEN Hui-Peng. Expression and Characterization of Fusion Protein of Interferon and Transferrin in Pichia pastoris[J]. Progress in Biochemistry and Biophysics,2005,32(7):625-629
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