生物工程學(xué)報(bào) 25 June 2009, 25(6):806～812
豬I型與II型干擾素的克隆、表達(dá)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(xué)院病原微生物及免疫學(xué)重點(diǎn)實(shí)驗(yàn)室 中國科學(xué)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(xì)胞受病毒感染或其他生物誘導(dǎo)劑刺激而產(chǎn)生的天然蛋白質(zhì), 主要功能為抗病毒增殖,、調(diào)節(jié)免疫反應(yīng)和激活免疫細(xì)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(gòu)建原核表達(dá)載體pET-His/PoIFN-a、pET-His/PoIFN-g,、pET-His/PoIFN- ag和pET-His/PoIFN-w, 轉(zhuǎn)化大腸桿菌Rosetta(DE3)進(jìn)行表達(dá), 經(jīng)純化,、復(fù)性得到具有生物學(xué)活性的蛋白。用細(xì)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(jìn)行抗病毒活性測定, 結(jié)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(dá)108 U/mg; 豬g干擾素活性效價(jià)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(jìn)一步驗(yàn)證,。本研究對(duì)干擾素在抗病毒、提高機(jī)體免疫方面的應(yīng)用提供了理論依據(jù),。
關(guān)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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