猪γ干扰素在家蚕中的表达和抗病毒活性检测  

Expression and Antiviral Activity Detection of Porcine Interferongamma in Silkworm

作　　者：刘兴健[1] 李皓洋 胡小元[1] 张志芳[1] 易咏竹[2] 李轶女[1] 

机构地区：[1]中国农业科学院生物技术研究所,北京100081 [2]中国农业科学院蚕业研究所,镇江212018

出　　处：《生物技术通报》2016年第1期144-148,共5页Biotechnology Bulletin

基　　金：国家高技术研究发展计划"863"(2011AA100603);国家重点基础研究发展计划(2012CB114600)

摘　　要：干扰素在畜牧养殖业中可以用于病毒性传染病的治疗和疫苗免疫效力的提高,用杆状病毒表达系统在家蚕中表达了猪γ干扰素。根据已发表的序列对猪γ干扰素基因的密码子进行优化并合成,将其克隆到杆状病毒转移载体p VL1393上,与Bm Bacmid病毒基因组DNA共转染Bm N细胞系,获得重组病毒,猪γ干扰素基因位于重组Bm NPV病毒的多角体基因启动子下游。用该重组病毒感染家蚕获得含有猪γ干扰素的表达产物。Western blotting检测到表达产物中的猪γ干扰素,用微量细胞病变法利用干扰素抑制VSV-GFP感染VERO细胞的方法来测定干扰素活性,结果显示干扰素效价可以达到6×105 IU/m L以上。在家蚕幼虫体内成功表达并获得了有活性的猪γ干扰素。Interferon is used for threating viral infectious diseases and enhancing vaccine efficacy in stockbreeding. In this study, recombinant porcine interferon-gamma（Po IFNγ）was expressed in silkworm by using Baculovirus expression vector system. Based on the published sequences, the gene of Po IFNγ was artificially synthesized after codon optimization. The optimized gene was firstly cloned into p VL1393 transfer vector of Baculovirus, with virus Bm Bacmid DNA which was co-transfected into Bm N cell line, and recombinant Bm NPV was obtained. The Po IFNγ gene was in the downstream of polyhedrosis gene promoter in Bm NPV. The silkworm larva infected with recombinant Bm NPV gained the expression of Po IFNγ that was detected by Western blotting. The activity of interferon was measured by using microcytopathic method that uses interferon to inhibit the VSV-GFP infecting VERO cells. The results showed that the titer of interferon was over 6×105 IU/m L. The activated Po IFNγ was successfully expressed in silkworm larvae.

关 键 词：猪Γ干扰素 家蚕 杆状病毒表达系统 抗病毒活性 

分 类 号：S859.79[农业科学—临床兽医学]