人表皮生长因子融合蛋白的表达及纯化工艺的优化

中国生物工程杂志

2010, Vol. 30

Issue (03): 74-78

技术与方法

人表皮生长因子融合蛋白的表达及纯化工艺的优化

张海淼^1,2,刘孝菊^1,3,田海山¹,万晓珊^1,2,邓林^1,2,李校堃^1,2,3*

1.吉林农业大学生物反应器与药物开发教育部工程研究中心长春 130118
2.吉林农业大学生命科学学院长春1301183 吉林大学公共卫生学院长春 1300021

Expression and Purification of Fusion Protein with SUMO of Human Epidermal Growth Factor

1.College of Life Sciences, Jilin Agricultural University, Changchun 130118, China
2.Engineering Research Center of Bioreactor and Pharmaceutical Development, Ministry of Education, Jilin Agricultural University，Changchun 130118, China

全文: PDF(681 KB) HTML

摘要：

探讨了在大肠杆菌中生产小分子泛素样修饰蛋白与人表皮生长因子（SUMO-hEGF）的最佳表达及纯化条件。将重组表达载体pET3c-SUMO-hEGF转化到大肠杆菌BL21(AI)中，以阿拉伯糖为诱导剂，对诱导表达参数进行优化，并进一步通过离子交换层析，Ni-NTA亲和层析及分子筛层析等进行纯化分析。结果表明： SUMO-hEGF在BL21(AI)中的最佳诱导表达温度为37℃，诱导剂阿拉伯糖的最佳浓度为5.0g/L，最佳诱导表达时间为4h，表达量约为20.2%，Western blot 分析证实，纯化后的蛋白是hEGF，为进一步开发hEGF基因工程药物奠定了基础。

关键词： 人表皮生长因子; 阿拉伯糖; 小分子泛素样修饰蛋白; 纯化

Abstract:

The condition of both expression and purification of SUMO human epidermal growth factor in E.coli is discussed. After transforming recombinant expression vector into E.coli, the experiment optimizes the induction expression, and analysis through ion exchange chromatography NiNTA affinity chromatography and molecular sieve chromatography, adopting arabinose as the IPTG. The results are as follows: 37℃ is the most favorable temperature for induction expression of SUMO-hEGF in BL21(AI), 5.0g/L the most best concentration of arabinose, and 4h the best time for induction expression. Meanwhile, the quantity of expression is around 20.2%. The conclusion that hEGF is the purified protein is verified by Western blot.The good foundation for the development of hEGF genetic medicine is founded.

Key words: Human Epidermal Growth Factor (hEGF) arabinose SUMO Purification

收稿日期: 2009-12-16 出版日期: 2010-03-25

通讯作者: 李校堃 E-mail: xiaokunli@163.com

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引用本文:

张海淼刘孝菊田海山万晓珊邓林李校堃. 人表皮生长因子融合蛋白的表达及纯化工艺的优化[J]. 中国生物工程杂志, 2010, 30(03): 74-78.

ZHANG Hai-Miao, LIU Xiao-Ju, TIAN Hai-Shan, MO Xiao-Shan, DENG Lin, LI Jiao-Kun. Expression and Purification of Fusion Protein with SUMO of Human Epidermal Growth Factor. China Biotechnology, 2010, 30(03): 74-78.

链接本文:

https://manu60.magtech.com.cn/biotech/CN/ 或 https://manu60.magtech.com.cn/biotech/CN/Y2010/V30/I03/74

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