生物工程學報  25 June 2009, 25(6):806～812

豬I型與II型干擾素的克隆,、表達及抗病毒活性比較

陳雪梅, 薛清華, 祝榮格, 付顯華, 楊利敏, 孫蕾, 劉文軍

中國科學院病原微生物及免疫學重點實驗室 中國科學院微生物研究所分子病毒中心, 北京 100101

摘  要: 干擾素(IFN)是由多種細胞受病毒感染或其他生物誘導劑刺激而產生的天然蛋白質, 主要功能為抗病毒增殖,、調節(jié)免疫反應和激活免疫細胞等。本研究克隆并測序了豬干擾素(PoIFN)a,、g,、a g及w基因。構建原核表達載體pET-His/PoIFN-a,、pET-His/PoIFN-g,、pET-His/PoIFN- ag和pET-His/PoIFN-w, 轉化大腸桿菌Rosetta(DE3)進行表達, 經純化、復性得到具有生物學活性的蛋白,。用細胞病變抑制法在Marc-145/PRRSV,、Marc-145/VSV、PK-15/VSV,、Vero/VSV,、MDBK/VSV系統(tǒng)上進行抗病毒活性測定, 結果表明豬a和ag融合干擾素有較為顯著的抗病毒活性, 抗PRRSV活性高達108 U/mg; 豬g干擾素活性效價約為a干擾素的1/2到1/3; 豬w干擾素幾乎未檢測到抗病毒活性, 需進一步驗證。本研究對干擾素在抗病毒,、提高機體免疫方面的應用提供了理論依據(jù),。

關鍵詞: 豬干擾素, 抗病毒活性

Comparison of antiviral activities of porcine interferon type I and type II

Xuemei Chen, Qinghua Xue, Rongge Zhu, Xianhua Fu, Limin Yang, Lei Sun, and Wenjun Liu

Chinese Academy of Sciences Key Laboratory of Pathogenic Microbiology and Immunology, Center for Molecular Virology, Institute of Microbiology, Chinese Academy of Sciences, Beijing 100101, China

Abstract: Interferons (IFNs) are natural proteins produced by wide variety of cells in response to viral infection or other biological inducers, and they execute diversified functions as antiviral defense, immune activation and cell growth regulation. Four genes encoding porcine interferons (PoIFN), PoIFN-α, PoIFN-γ, PoIFN-αγ or PoIFN-ω, were cloned and sequenced. The four types of porcine interferon genes were subcloned into the pET-His vector, and expressed in Escherichia coli Rosetta (DE3). The recombinant products were purified and renaturalized from inclusion bodies to obtain a native state of well biological activity. Antiviral activity assays for porcine interferons were performed and evaluated by standard procedures in following cell/virus test systems: Marc-145/PRRSV, Marc-145/VSV, PK-15/VSV, Vero/VSV or MDBK/VSV. The data showed that both PoIFN-α and PoIFN-αγ demonstrated significant antiviral activities, and the titer of them against PRRSV was up to 108 U/mg. PoIFN-γ had approximately half or one-thirds antiviral activity of PoIFN-α. PoIFN-ω showed inconspicuous antiviral activity.

Keywords: porcine interferon, antiviral activity

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