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当前位置: 首页 > 文章 > 重组鸡γ-干扰素在家蚕中的表达及抗病毒活性 蚕业科学 2018 (6) 898-904
Position: Home > Articles > Expression of Recombinant Chicken Interferon-γ in Bombyx mori with Baculovirus Expression System and Determination of Its Antiviral Activity Science of Sericulture 2018 (6) 898-904

重组鸡γ-干扰素在家蚕中的表达及抗病毒活性

作  者：
赵泽;赵璐璐;刘兴健;王朋;李轶女;张志芳;易咏竹
单  位：
江苏科技大学生物技术学院;中国农业科学院生物技术研究所
关键词：
鸡γ-干扰素;杆状病毒表达系统;家蚕;抗病毒活性
摘  要：
鸡γ-干扰素(Ch IFN-γ)是一类具有免疫调节及抗病毒活性的细胞因子。利用家蚕杆状病毒表达系统表达具有高生物活性的鸡γ-干扰素。首先,对Ch IFN-γ基因序列进行优化与合成,再将其克隆到杆状病毒转移载体p VL1393多克隆位点中,与缺少orf1629基因的家蚕杆状病毒re Bm Bac共转染Bm N细胞获得重组病毒re Bm-ChIFN-γ。将re Bm-ChIFN-γ注射感染家蚕5龄幼虫,检测感染后96～120 h之间发病蚕体的血淋巴。PCR扩增结果显示成功构建了重组杆状病毒re Bm-ChIFN-γ,Western blot检测重组杆状病毒在家蚕体内表达了目的干扰素,微量细胞病变抑制法检测表达产物具有明显的抗病毒活性,效价约为(3. 24±0. 32)×10~6U/mL。试验结果表明,利用家蚕杆状病毒表达系统成功地表达出具有较高抗病毒活性的鸡γ-干扰素,为鸡γ-干扰素相关生物制剂的研发奠定了基础。
译  名：
Expression of Recombinant Chicken Interferon-γ in Bombyx mori with Baculovirus Expression System and Determination of Its Antiviral Activity
作  者：
Zhao Ze;Zhao Lulu;Liu Xingjian;Wang Peng;Li Yi'nyu;Zhang Zhifang;Yi Yongzhu;College of Biotechnology,Jiangsu University of Science and Technology;Biotechnology Research Institute,Chinese Academy of Agricultural Sciences;
单  位：
Zhao Ze%Zhao Lulu%Liu Xingjian%Wang Peng%Li Yi'nyu%Zhang Zhifang%Yi Yongzhu%College of Biotechnology,Jiangsu University of Science and Technology%Biotechnology Research Institute,Chinese Academy of Agricultural Sciences
关键词：
Chicken interferon-γ;;Baculovirus expression system;;Silkworm;;Antiviral activity
摘  要：
Chicken gamma interferon( Ch IFN-γ) is a kind of cytokine which plays a role in immune regulation and antiviral activity. Here,we expressed Ch IFN-γ with high biological activity by silkworm baculovirus expression system.First of all,the codons of Ch IFN-γ were optimized and the gene was synthesized and cloned into multiple cloning site of baculovirus transfer vector pVL1393. Then, we co-transfected Bm N cells with both the recombinant plasmid and re Bm Bac bacmid without orf1629 gene,obtained and purified the recombinant virus re Bm-ChIFN-γ. Subsequently,we injected 5 th instar larvae of Bombyx mori with re Bm-ChIFN-γ,collected the hemolymph of silkworm 96-120 h after injection and preserved for further detection. PCR results suggested that we constructed the recombinant baculovirus re BmCh IFN-γ successfully and target protein expression was confirmed by Western blot. In addition,biological potency of the protein was approximately( 3. 24± 0. 32) × 10~6 U/mL as determined by cytopathic effect inhibition assay. We conclude that,the Ch IFN-γ with high biological activity was expressed successfully in silkworm bioreactor through silkworm baculovirus expression system,which sets the good stage for subsequent biological agents related to Ch IFN-γ.

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