| 植物甜蛋白des-pGlu1-Brazzein的密码子优化及其在毕氏酵母中的表达 |
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| 引用本文： | 李春丽,韩露,郑振宇,赵卫东.植物甜蛋白des-pGlu1-Brazzein的密码子优化及其在毕氏酵母中的表达[J].生物工程学报,2011,27(8):1158-1163. |
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| 作者姓名： | 李春丽 韩露 郑振宇 赵卫东 |
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| 作者单位： | 河南农业大学牧医工程学院,郑州,450002 |
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| 基金项目： | 河南省教育厅基础研究项目 (No. 2004922043)，河南农业大学博士基金项目资助。 |
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| 摘 要： | 根据甜蛋白des-pGlu1-Brazzein的成熟区氨基酸序列，按照酵母密码子的偏好优化其编码序列，合成了4对末端具粘合位点的寡聚核苷酸序列，经连接、PCR扩增后得到了179 bp的des-pGlu1-Brazzein编码序列，插入到pPIC9K载体中，构建了重组表达载体pPIC9K-Bra。经酶切、电击转化到毕赤酵母菌GS115中。酵母工程菌株经筛选鉴定后诱导表达，目的蛋白的表达量约占上清总蛋白的51.6％，分离纯化后的des-pGlu1-Brazzein具有一定的甜度。
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| 关 键 词： | 甜蛋白，毕氏酵母，表达，活性检测 |
| 收稿时间： | 2010/12/2 0:00:00 |
Optimization of plant des-pGlu1-Brazzein gene according to yeasty biased codons and its expression in Pichia pastoris |
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| Chunli Li,Lu Han,Zhenyu Zheng and Weidong Zhao.Optimization of plant des-pGlu1-Brazzein gene according to yeasty biased codons and its expression in Pichia pastoris[J].Chinese Journal of Biotechnology,2011,27(8):1158-1163. |
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| Authors: | Chunli Li Lu Han Zhenyu Zheng and Weidong Zhao |
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| Institution: | College of Animal Science and Veterinary Medicine, Henan Agricultural University, Zhengzhou 450002, China;College of Animal Science and Veterinary Medicine, Henan Agricultural University, Zhengzhou 450002, China;College of Animal Science and Veterinary Medicine, Henan Agricultural University, Zhengzhou 450002, China;College of Animal Science and Veterinary Medicine, Henan Agricultural University, Zhengzhou 450002, China |
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| Abstract: | According to the amino acid sequence of des-pGlu1-Brazzein, 4 pairs of oligonucleotide with cosmic site were synthesized by using yeasty biased codons. After linkage and PCR, the 179 bp code area of des-pGlu1-Brazzein was obtained and inserted into pPIC9K, which resulted in the recombinant expression vector pPIC9K-Bra. By digestion with Sal I, the lined pPIC9K-Bra was transformed into Pichia pastoris GS115 by electric shock. The results of expression indicted that the secreted target protein accounted for 51.6% of total protein in the supernatant and showed biological activity after purification. |
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| Keywords: | des-pGlu1-Brazzein Pichia pastoris expression biological activity |
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