| 甜味蛋白(brazzein)基因的人工合成及其在大肠杆菌中的重组表达 |
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| 引用本文： | 刘松财,张永亮,任晓慧,张明军,柳巨雄,冯立文.甜味蛋白(brazzein)基因的人工合成及其在大肠杆菌中的重组表达[J].中国兽医学报,2006,26(5):538-540. |
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| 作者姓名： | 刘松财 张永亮 任晓慧 张明军 柳巨雄 冯立文 |
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| 作者单位： | 吉林大学,畜牧兽医学院,吉林,长春,130062 |
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| 摘 要： | 根据从西非植物Pentadiplandra brazzeana Baillion果实中分离的甜味蛋白brazzein的成熟区氨基酸序列，采用大肠杆菌（Escherichia coli）偏爱的密码子，人工合成了5对寡聚核苷酸序列，经体外连接和PCR扩增后得到了187bp的brazzein的编码序列，将该序列插入pET-28a载体构建成重组表达载体pET-Br。将pET-Br转化至大肠杆菌BL-21细胞，诱导表达后得到了与预计相对分子质量相同的蛋白。该蛋白约占细菌可溶性蛋白的18．9％，分离纯化后通过蛋白复性，得到有微甜味的产物。
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| 关 键 词： | 甜味蛋白 重组表达 |
| 文章编号： | 1005-4545(2006)05-0538-03 |
| 收稿时间： | 2005-07-11 |
| 修稿时间： | 2005年7月11日 |
Synthesis of Brazzein Gene and Its Recombinant Expression in Escherichia coli |
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| LIU Song-cai,ZHANG Yong-liang,REN Xiao-hui,ZHANG Ming-jun,LIU Ju-xiong,FENG Li-wen.Synthesis of Brazzein Gene and Its Recombinant Expression in Escherichia coli[J].Chinese Journal of Veterinary Science,2006,26(5):538-540. |
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| Authors: | LIU Song-cai ZHANG Yong-liang REN Xiao-hui ZHANG Ming-jun LIU Ju-xiong FENG Li-wen |
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| Institution: | College of Animal Science and Veterinary Medicine,Jilin University,Changchun 130062,China |
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| Abstract: | According to the amino acid sequence of Brazzein isolated from the fruit of Pentadiplandra baillion ,five pairs of oligonucleotides were synthesized by using Escherichia coli biased codons. After ligating and PCR amplification, 187 bp coding region of brazzein was obtained and inserted into plasmid pET-28a. The results of induced expression showed that the molecular weight of the protein was the same as expected,accounting for 18. 9% of total soluble protein of the E. coli. The protein after being denatured and renatured bears a lightly sweet taste. |
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| Keywords: | brazzein |
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