| 甜蛋白Monellin基因在大肠杆菌中的高效表达 |
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| 引用本文： | 陈忠军,蔡恒,路福平,杜连祥.甜蛋白Monellin基因在大肠杆菌中的高效表达[J].生物工程学报,2005,21(4):568-572. |
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| 作者姓名： | 陈忠军 蔡恒 路福平 杜连祥 |
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| 作者单位： | 天津市工业微生物重点实验室,天津科技大学生物工程学院,天津,300222 |
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| 摘 要： | 据已报道的单链monellin甜蛋白的氨基酸序列，采用细菌偏爱密码子，人工合成了全长 294bp的 monellin基因。插入到大肠杆菌表达载体Pet_22b中，构建重组分泌型表达载体Petmo。经IPTG诱导Petmo所含有的甜蛋白基因可在大肠杆菌BL21（DE3）中高效表达，表达量占菌体可溶性蛋白的44.8％。且经纯化后测定其甜度是蔗糖的3000倍。得到的甜蛋白热稳性及耐酸性均比天然产物有所提高。
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| 关 键 词： | 甜蛋白Monellin 细菌优化密码子 重组PCR 诱导表达 |
| 文章编号： | 1000-3061(2005)04-0568-05 |
| 修稿时间： | 2004年10月1日 |
Overexpression of a Sweet Protein Monellin in Escherichia coli |
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| CHEN Zhong-Jun,CAI Heng,LU Fu-ping,DU Lian-xiang.Overexpression of a Sweet Protein Monellin in Escherichia coli[J].Chinese Journal of Biotechnology,2005,21(4):568-572. |
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| Authors: | CHEN Zhong-Jun CAI Heng LU Fu-ping DU Lian-xiang |
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| Institution: | Tan Key Lab of Industrial Microbiology, The College of Biotechnology, Tianjin University of Science & Technology, Tianjin 300222, China. |
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| Abstract: | According to the amino acid sequence of monellin, a single chain 294bp monellin gene was synthesized and inserted into vector pET-22b to yield the recombinant secretion plasmid pETMO. The single-chain monellin gene was designed based on the biased codons of E.coli so that its expression would be then optimized. Under the expressing conditions, monellin was produced accounting for 44.8% of total soluble proteins. The E.coli-expressed single-chain monellin is 3000 times sweeter than sucrose. The thermal-stability and acid-resistance of the protein are higher than the natural monellin. |
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| Keywords: | monellin bacteria preferred condon recombination PCR inducing expression |
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