重组猪γ-干扰素的表达与纯化  

Expression and purification of recombinant porcine interferon-γ

作　　者：王荣谈 郭佳宏[2] 彭丽英[2] 廖学文[2] 姚慧娟[2] 樊生超[2] 缪德年[2] 

机构地区：[1]上海瑞丰农业科技有限公司,上海201106 [2]上海市农业科学院,上海201106

出　　处：《上海农业学报》2014年第2期10-14,共5页Acta Agriculturae Shanghai

基　　金：上海市科技支撑项目(13391901500)资助

摘　　要：将猪γ-干扰素（Porcine interferon-γ，PolFN-γ）成熟蛋白的基因合成克隆到表达载体pET30a中，转化大肠杆菌BL21（DE3）中表达。对重组菌的表达条件进行优化，发现在IPTG体积浓度为0．04mmol／L、诱导时间为3h、诱导温度为30℃的条件下，目的蛋白大部分以可溶形式表达。重组蛋白经亲和柱纯化后的纯度大于90％，表达量达40mg／L菌液。重组蛋白在MDBK细胞上抗滤泡性口炎病毒的活性为2×10^6U／mg。A synthetic gene of porcine interferon-γ （PoIFN-γ）mature protein was inserted into vector pET30a and expressed in E. coil BL21 （DE3）strain. The optimization of the recombinant strain expression conditions showed that most of the target protein was expressed in a soluble form under the conditions of 0.04 mmol/L IPTG（isopropyl-β-D-thiogalactoside）and 3 h induction at 30 ℃. Through affinity column purification the recombinant protein was greater than 90% in purity and the expression level of culture was up to 40 mg/L. The antiviral activity of the recombinant protein was about 2 × 10^6 U/mg in the MDBK/VSV system test.

关 键 词：猪Γ-干扰素 可溶性表达 纯化 

分 类 号：S852.65[农业科学—基础兽医学]