| 荧光素酶基因luc在大肠杆菌中表达条件优化 |
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| 引用本文： | 夏蕾,饶志明,沈微,方慧英,诸葛健.荧光素酶基因luc在大肠杆菌中表达条件优化[J].生物学杂志,2009,26(4):5-8. |
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| 作者姓名： | 夏蕾 饶志明 沈微 方慧英 诸葛健 |
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| 作者单位： | 江南大学工业生物技术教育部重点实验室江南大学工业微生物研究中心,江苏,无锡,214122 |
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| 基金项目： | 国家自然科学基金，国家863高科技研究发展计划，江苏省青年科技创新人才基金，长江学者和创新团队发展计划资助 |
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| 摘 要： | 对重组荧光素酶大肠杆菌菌株M15／pQE30-luc进行了表达条件的优化研究。单因素结果表明：在初始pH值7．0，装液量为20％，2％的接种量，终浓度为0．5mmol／L的IPTG，添加10—30mmol／L的Mg^2＋，摇床转速为200r／min，37℃诱导3．5h酶的表达量最高。正交试验结果表明：初始pH值为7．0，添加40mmol／LMg^2=，接种量2％，装液量为20％时表达量最高，比酶活达1．63×10^8RFU／mg蛋白。
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| 关 键 词： | 萤火虫荧光素酶 表达 发酵条件 正交试验设计 |
Optimization of the expression of luc gene in E. coil |
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| XIA Lei,RAO Zhi-ming,SHEN Wei,FANG Hui-ying,ZHUGE Jian.Optimization of the expression of luc gene in E. coil[J].Journal of Biology,2009,26(4):5-8. |
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| Authors: | XIA Lei RAO Zhi-ming SHEN Wei FANG Hui-ying ZHUGE Jian |
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| Institution: | ( Key Laboratory of Industrial Biotechnology, Ministry of Education, Research Center of Industrial Microorganisms, Jiangnan University, Wuxi 214122, China) |
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| Abstract: | The luc gene was subcloned into expression vector pQE30 and its product LUC was overexpressed in Escherichia coli M15. The optimum fermentation condition was： pH 7.0, the amount of medium 20% , inoculum size 2%, the final concentration of IPTG 0.5 mmoL/L, 10 - 30 mmol/L Mg^2 ＋ , the rotating speed 200 r/min, inducing 3.5h at 37℃. After orthogonal experimental design, the result showed that the initial pH7.0, 40mmol/L Mg^2＋ , 2% inoculation amount and the 20% medium amount had the best expression of LUC, and the specific activity was 1.63 × 10^8 RFU/mg protein. |
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| Keywords: | firefly luciferase expression fermentation condition orthogonal experimental design |
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