SONG Jing,ZHU Jianquan,ZHANG Quanbao,et al.Optimization of Prokaryotic Expression of Recombinant Caffeic Acid 3-O-methyltransferase From Ligusticum chuanxiong in E.coli[J].Northern Horticulture,2016,40(18):107-110.[doi:10.11937/bfyy.201618026]
川芎咖啡酸-3-O-甲基转移酶的原核表达与条件优化
- Title:
- Optimization of Prokaryotic Expression of Recombinant Caffeic Acid 3-O-methyltransferase From Ligusticum chuanxiong in E.coli
- 关键词:
- 川芎; 咖啡酸-3-O-甲基转移酶; 原核表达; 条件优化
- Keywords:
- Ligusticum chuanxiong; caffeic acid 3-O-methyltransferase; prokaryotic expression; conditions optimization
- 文献标志码:
- A
- 摘要:
- 以川芎咖啡酸-3-O-甲基转移酶基因（LCCOMT）为表达对象，将含有6*His标签的LCCOMT基因连接pET28a表达载体后转化大肠杆菌BL21，采用原核表达的方法，优化了LCCOMT基因的表达条件。结果表明：当诱导时间为4 h，IPTG终浓度为20 mmol?L-1，诱导温度为37 ℃时，重组LCCOMT蛋白的表达量达到最大。溶解性检测结果表明，LCCOMT重组蛋白以可溶性蛋白的形式存在，占总蛋白的9%，该试验可为LCCOMT的大规模表达纯化奠定基础。
- Abstract:
- In order to efficiently express Ligusticum chuanxiong caffeic acid 3-O-methyltransferase (LCCOMT) in E.coli，pET28a-LCCOMT expression vector，which contained LCCOMT gene with 6*His labels，was transformed into E.coli BL21.The expression conditions of LCCOMT gene were optimized.The results showed that when the induction time was 4 hours，IPTG concentration was 20 mmol?L-1 and temperature was 37 ℃，the expression of LCCOMT recombinant protein might reach the maximum.Furthermore，solubility test results showed that the LCCOMT recombinant protein existed in the form of soluble protein.Therefore，this research laid a foundation for the future large-scale expression and purification of LCCOMT protein.
参考文献/References:
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备注/Memo
第一作者简介：宋婧（1995-），女，本科，研究方向为生物学。E-mail:1344366035@qq.com.责任作者：周嘉裕(1976-),女，博士，副教授，研究方向为药用植物分子生物学。E-mail：spinezhou@home.swjtu.edu.cn.基金项目：国家自然科学基金资助项目（31371232，31500276）；成都市科技技术研发资助项目（2015-HM01-00051-SF）；国家级大学生创新创业训练计划资助项目（201510613063）。