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| 人乙醛脱氢酶2的原核表达及其条件优化 | |
| 引用本文： | 孙莉,李智博,秦飞,邓帅,杨君.人乙醛脱氢酶2的原核表达及其条件优化[J].基因组学与应用生物学,2012,31(2):123-128. |
| 作者姓名： | 孙莉 李智博 秦飞 邓帅 杨君 |
| 作者单位： | 1. 大连海洋大学,食品科学与工程学院,大连,116023;大连理工大学,生命科学与技术学院,大连,116024 2. 大连海洋大学,食品科学与工程学院,大连,116023 3. 大连理工大学,生命科学与技术学院,大连,116024 |
| 基金项目： | 中央高校基本科研业务费专项资金 |
| 摘 要： | 为实现人乙醛脱氢酶2(ALDH2)基因在原核生物中高效表达,将含有6×His标签和SUMO融合蛋白标签的人乙醛脱氢酶2基因的表达载体转化至宿主菌BL21(DE3)中。在异丙基硫代-β-D-半乳糖苷(IPTG)诱导下,目的基因在大肠杆菌内高效表达。通过对表达条件的优化,37℃使用终浓度0.3mmol/L的IPTG诱导3h,重组大肠杆菌的表达量可占全菌蛋白的16%。SUMO融合蛋白标签的加入以及较低的诱导温度(16℃)有利于提高人乙醛脱氢酶2基因在大肠杆菌内的可溶性表达。 |
| 关 键 词： | 乙醛脱氢酶 原核表达 SUMO蛋白 表达优化 |
Expression and Optimization of Human Aldehyde Dehydrogenase 2 in Escherichia coli |
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| Sun Li , Li Zhibo , Qin Fei , Deng Shuai , Yang Jun.Expression and Optimization of Human Aldehyde Dehydrogenase 2 in Escherichia coli[J].Genomics and Applied Biology,2012,31(2):123-128. | |
| Authors: | Sun Li Li Zhibo Qin Fei Deng Shuai Yang Jun |
| Institution: | 1 College of Food Science and Engineering,Dalian Ocean University,Dalian,116023;2 School of Life Science and Biotechnology,Dalian University of Technology,Dalian,116024 * Corresponding author,zhibo1979@163.com DOI:10.3969/gab.031.000123 |
| Abstract: | In order to highly express the human aldehyde dehydrogenase 2(ALDH2) gene in prokaryotic cells,ALDH2 gene was subcloned into the expression vector with N-terminal SUMO-6his-tag,then the recombinant plasmid was transformed into BL21(DE3).With the inducement of IPTG,the recombinant human ALDH2 gene was expressed efficiently.Through optimizing the conditions for expression(0.3 mmol/L IPTG,37℃,3 h),the amount of the expression of the recombinant human ALDH2 achieved about 16% of total protein in culture medium.The adding of SUMO tag and the moderately low induction temperature can improve the soluble expression of human ALDH2. |
| Keywords: | Aldehyde dehydrogenase Prokaryotic expression SUMO tag Optimal expression conditions |
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