| 短短小芽孢杆菌大肠杆菌穿梭分泌表达载体的构建 |
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| 引用本文： | 彭清忠,张惟材,朱厚础.短短小芽孢杆菌大肠杆菌穿梭分泌表达载体的构建[J].生物工程学报,2002,18(4):438-441. |
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| 作者姓名： | 彭清忠 张惟材 朱厚础 |
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| 作者单位： | 军事医学科学院生物工程研究所,北京,100071 |
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| 基金项目： | 军事医学科学院创新课题启动基金资助课题 (No .0 0 10 0 18)~~ |
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| 摘 要： | 应用PCR技术从具有分泌蛋白能力强且没有胞外蛋白酶活性的短短小芽孢杆菌50中分离出细胞壁蛋白基因的多启动子和信号肽编码序列，利用它与质粒pUB110和pKF3-起构建成穿梭分泌表达载体pBKE50，将α0淀粉酶基因引入该载体转化短短小芽孢杆菌50后，发现α－淀粉酶可以活性形式分泌表达，此工作为下一步建立短短小芽孢杆菌高效分泌表达系统奠定了基础。
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| 关 键 词： | 短短小芽孢杆菌－大肠杆菌 穿梭分泌表达载体 构建 |
| 文章编号： | 1000-3061(2002)04-0438-04 |
| 修稿时间： | 2002年1月25日 |
The Construction of Shuttle Vectors of Brevibacillus brevis-Escherichia coli |
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| PENG Qing-Zhong\ ZHANG Wei-Cai\ ZHU Hou-Chu\.The Construction of Shuttle Vectors of Brevibacillus brevis-Escherichia coli[J].Chinese Journal of Biotechnology,2002,18(4):438-441. |
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| Authors: | PENG Qing-Zhong\ ZHANG Wei-Cai\ ZHU Hou-Chu\ |
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| Institution: | Institute of Biotechnology, Academy of Military Medical Sciences, Beijing 100071, China. |
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| Abstract: | The 5' region of the cell wall protein(CWP) gene containing multiple tandem promoters and the signal peptide-coding sequence was isolated by PCR from Br. brevis 50, and used to construct the shuttle vector pBKE50, which included the replication origin of pUB110 and the erythromycin-resistance gene of pGK12. The alpha-amylase gene of Bacillus subtilis 168 was ligated to pBKE50, producing plasmid pBKE50/alpha-amy. After the resulting plasmid was introduced into Br. brevis 50, soluble and biologically active alpha-amylase was secreted directly into the culture medium. The expression level of alpha-amylase in the recombinant Br. brevis 50 was twice higher than that of the donor strain. |
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| Keywords: | Brevibacillus brevis shuttle vector secretory expression |
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