Curr. Biotech. ›› 2013, Vol. 3 ›› Issue (4): 252-256.DOI: 10.3969/j.issn.2095-2341.2013.04.05

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Expression of Porcine Interferon-alpha in Silkworm, Bombyx mori and its Antiviral Activity Detection

ZHONG Lu-long, YI Yong-zhu, DING Nong, ZHANG Jin-wei, LIU Ji-xing   

  1. 1.State Key Laboratory of Veterinary Etiological Biology; Key Laboratory of Grazing Animal Dieases,  Ministry of Agriculture; Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou 730046, China;
    2.Sericultural Research Institute, Chinese Academy of Agricultural Sciences, Jiangsu Zhenjiang 212018, China;
    3.Huzhou Academy of Agricultural Sciences, Zhejiang Huzhou 313000, China
  • Received:2013-05-17 Online:2013-07-25 Published:2013-06-14

猪α干扰素在家蚕中表达及其抗病毒活性测定

钟鲁龙1,易咏竹2,丁农3,张金卫3,柳纪省1*   

  1. 1.中国农业科学院兰州兽医研究所, 家畜疫病病原生物学国家重点实验室, 农业部草食动物疫病重点开放实验室, 兰州 730046;
    2.中国农业科学院蚕业研究所, 江苏 镇江 212018;
    3.湖州市农业科学院, 浙江 湖州 313000
  • 通讯作者: 柳纪省,研究员，从事动物传染病及其病原分子流行病学研究。Tel:0931-8342682;E-mail:liujixing@hotmail.com
  • 作者简介:钟鲁龙,硕士研究生，研究方向为动物传染病及其病原分子流行病学。E-mail:longginger@163.com。
  • 基金资助:

    甘肃省高层次人才科技创新创业扶持行动项目(1013JHTA008)资助。

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Abstract: Porcine interferon-alpha(PoIFNα) was expressed in silkworm larvae using silkworm-baculovirus expression system. The porcine interferon-alpha gene was artificially synthesized after codon optimization. The optimized target gene was cloned to the baculovirus transfer vector pVL1393 and the recombinant plasmid pVL-PoIFNα was constructed. pVL-PoIFNα was co-transfected with linearized Bm-BacPAK6 DNA into Bm-N cells. The recombinant proteins were successfully detected in hemolymph by Western blot analysis. The recombinant PoIFN-alpha expressed in hemolymph were verified to be of high antiviral activity by inhibiting the cytopathic effect of VSV*GFP in Vero cells, which was about 106 U/mL.

Key words: porcine interferon, antiviral activity, silkworm, baculovirus, eukaryotic expression

摘要： 本研究利用家蚕杆状病毒表达系统在家蚕中表达猪α干扰素。首先优化并合成猪α干扰素基因，将其克隆到杆状病毒转移载体pVL1393的多角体基因启动子下游，与线性化的Bm-BacPAK6 DNA共转染至Bm-N细胞，在细胞内进行同源重组后，纯化得到重组病毒。Western杂交结果显示，在蚕血淋巴中可检测到猪α干扰素表达。用微量细胞病变抑制法在Vero/VSV*GFP系统上进行干扰素活性测定，结果表明重组猪α干扰素有抗病毒活性，效价达到106 U/mL以上，证明在家蚕幼虫体内成功表达了有活性的猪α干扰素。

关键词: 猪干扰素, 抗病毒活性, 家蚕, 杆状病毒, 真核表达

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