A loop-controlled rrnB P1 promoter for high-level expression of heterologous proteins in Escherichia coli

A loop-controlled rrnB P1 promoter for high-level expression of heterologous proteins in Escherichia coli
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用于在大肠杆菌中高水平表达异源蛋白的环控rrnB P1启动子

DOI:
10.1007/s10529-010-0426-2
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发表时间:
2011-02
影响因子:
2.7
通讯作者:
--
中科院分区:
工程技术4区
文献类型:
--
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利用rRNA rrnB P1启动子和lac操纵子调控元件(lacO)在大肠杆菌中构建了一个有效的蛋白表达体系。为了调节rrnB P1启动子的转录活性,我们设计了两个lacO位点,它们具有中间环结构;通过测定β-1,4-葡聚糖酶基因cel5G的表达水平来验证表达。与T7启动子相比,环状启动子结构的基础表达减少了92%。我们还发现宿主细胞类型对rrnB P1启动子的调控有显著影响:大肠杆菌DH5α和DH10B表达水平较高,而在BL21(DE3)中的表达更为严格。
An effective protein expression system was constructed in Escherichia coli utilizing the rRNA rrnB P1 promoter and the regulatory element of the lac operon (lacO). To regulate the transcriptional activity of the rrnB P1 promoter, we designed two lacO sites with an intervening loop structure; expression was verified by measuring the levels of the β-1,4-glucanase gene, cel5G. Basal expression from the looped promoter construct was reduced by 92% when compared to expression from the T7 promoter. We also found that the host cell type had a significant effect on the regulation of the rrnB P1 promoter: E. coli DH5α and DH10B had high expression levels, whereas the expression in BL21(DE3) was more stringent.
DOI: 10.1126/science.274.5294.1930
发表时间: 1996-12-13
期刊: SCIENCE
影响因子: 56.9
作者:
Fried, MG;Hudson, JM
通讯作者: Hudson, JM
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