Expression of Chicken Interferon-β in Pichia pastoris and Its Antiviral Effect |
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Authors: | LIU Xin-wen WANG Xiu-li GUO Dong-chun GUO Wei-wei ZOU Min FAN Gen-cheng |
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Institution: | 1. The Qingdao Yebio Biological Engineering Co., Ltd., Qingdao 266032, China;
2. Harbin Veterinary Research Institute, Haerbin 150069, China |
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Abstract: | In order to study the expression of chicken interferon-β (ChIFNβ) in Pichia pastoris and its antiviral effect,ChIFNβ gene was subcloned into pPICZα-A vector and constructed pPIC-ChIFNβ recombinant plasmid. After linearized by Sac Ⅰ, the plasmid was transferred into Pichia pastoris X-33 strain. The recombinant strain was induced by 1.0% methanol for 4 d,the product of recombinant ChIFNβ was detected by SDS-PAGE,the antiviral activity was determined by classical cell pathological effect inhibition assay. The protein weight was approximate 17 ku and the recombinant protein was secreted effectively in Pichia pastoris. Potency of interferon was 107.5 U/mL. The expressed protein could also inhibit H9N2 avian influenza virus proliferation in chick embryo. Animal experiment suggested that recombinant protein could provide 70% immune protection for aviadenovirus. The results showed that the recombinant ChIFNβ had good antivirus activity,it would be reference for clinical application of ChIFNβ. |
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Keywords: | chicken interferon-β secreted expression antiviral activity |
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