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作 者:杨光[1] 王文静[2] 徐良[1] 杨好冬[1] 王小纯[1]
机构地区:[1]河南农业大学生命科学学院,河南郑州450002 [2]郑州牧业工程高等专科学校,河南郑州450011
出 处:《郑州牧业工程高等专科学校学报》2013年第2期4-8,共5页Journal of Zhengzhou College of Animal Husbandry Engineering
基 金:河南省科技攻关项目(102101310600)
摘 要:为了提高大肠杆菌中重组Exendin-4的表达量,给中试奠定基础,利用单因素试验和正交试验研究了培养基、接种量、诱导时机、诱导温度、诱导剂浓度、诱导时长以及MgSO4浓度等对重组大肠杆菌生长和融合蛋白表达的影响。结果表明最佳培养基为2×YT,最佳接种量为3%,于细菌对数生长期中后期(OD600约1.5)开始诱导,诱导温度为34℃、乳糖浓度为0.1g/L,于诱导9h后重组Exendin-4表达量最大。In order to improve the amount of recombinant Exendin--4 expression in Escherichia coli and lay a foundation for the fermentation experiment. The effects of medium, inoculum size, the induction start--up time, temperature, the lactose concentration, the post--induction time and the concentration of MgSO4 on the growth of E. coli and the expression of the fusion protein were studied with single factor and orthogonal experiments. The results showed that the optimum culture medium was 2XYT and the optimum inoculum size was 3%. Adding lactose of 0. 1 g/L to the medium in the middle or later logarithmic growth phase (when OD600 reached approximately 1.5) and inducing at 34 ℃ for 9 h,the expression of recombinant Exendin--4 reached the highest level.
关 键 词:重组Exendin-4 工程菌 优化 表达
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