重组鸭α-干扰素基因工程菌表达条件的研究  

Studies on the Expression Conditions of Engineering E.coli Expressing Recombinant Duck Interferon-α

作　　者：杨梅[1] 程安春[1] 汪铭书[1] 张瑶[2] 龚永强[1] 陈斌[1] 

机构地区：[1]四川农业大学动物科技学,四川雅安625014 [2]动物疫病与人类健康四川省重点实验室,四川雅安625014

出　　处：《四川农业大学学报》2007年第3期337-342,共6页Journal of Sichuan Agricultural University

基　　金：国家科技攻关重大项目(2004BA901A03);教育部"新世纪优秀人才支持计划"项目(NCET-04-0906);四川省教育厅重点项目(2003A024);四川省重点建设学科项目(SZD0418)。

摘　　要：摘要：为了确定重组鸭α-干扰素（DuIFN—α）基因工程菌BL21（DE3）/pEW32a^＋-DuIFN—α的最佳表达条件，对影响原核表达的主要因素：IPTG浓度、诱导时期、收获时间、培养诱导温度、葡萄糖浓度及培养基进行了优化；并利用抗性筛选对表达质粒的稳定性进行了研究。结果显示，诱导基因工程菌BL21（DE3）/pEW32a^＋ -DulFN—a表达DuIFN—α的最佳条件为：以TB＋5g九葡萄糖培养基37℃培养至菌体OD600=0．8～1．1时，加入IPTG至终浓度0．2mmol/L，30℃诱导培养4～5h。在最佳表达条件下，DuIFN—α的相对含量为34．1％，总含量（终菌体浓度×相对含量）达79．9；各种优化条件对表达质粒的稳定性未见明显影响。To determine the optimal condition of the genetic engineering E. coli BL21 （DE3）/ pET32a^＋ - DulFN - α which expressed duck interferon -α, IPTG concentration, induction time, harvest period, cultivation temperatures, glucose concentration and culture medium were optimized in this study and plasmid stability was studied by the screening of resistance. The optimal inductive conditions were as follows： the culture medium was TB ＋ 5 g/L glucose, the culture temperature and the induction temperature were 37 ℃ and 30 ℃, OD600 was 0.8 -- 1.1, the concentration of IPTG was 0.2 mmol/L and the induction time was 4-5 h. By optimizing way, the relative amount of duck interferon - α was 34.1%, but there was no signal difference on plasmid stability by screening of resistance.

关 键 词：重组鸭α-干扰素 表达条件 优化 

分 类 号：S834[农业科学—畜牧学] S852.4[农业科学—畜牧兽医]