猪α干扰素在家蚕中表达及其抗病毒活性测定

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

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-BacPAK6DNA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106U/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。

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-transfeeted with lineafized Bm-BaePAK6 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 eytopathic effect of VSV ＊ GFP in Vero cells, which was about 106 U/mL.