Far different levels of gene expression provided by an oriented cloning system in Bacillus subtilis and Escherichia coli
Far different levels of gene expression provided by an oriented cloning system in Bacillus subtilis and Escherichia coli
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DOI：
10.1016/s0378-1097(03)00171-x
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发表时间：
2003-04-11
影响因子：
2.1
通讯作者：
Itaya, M
中科院分区：
文献类型：
--
作者：
Ohashi, Y;Ohshima, H;Itaya, M
建立了枯草芽孢杆菌和大肠杆菌的基因表达体系。表达载体pHASH102产生启动子和开放阅读框的任意组合，基于t扩展克隆法进行表达。由于pHASH系列载体被设计为在枯草芽孢杆菌基因组和高拷贝质粒之间穿梭，因此可以系统地检查拷贝数依赖性的表达谱。我们证明了带有Pr、Pspac和PS10启动子的载体适合于GFPuv的过表达。此外，编码枯草芽孢杆菌链霉素抗性基因氨基糖苷6-腺苷基转移酶的aadK在枯草芽孢杆菌和大肠杆菌中均成功过表达。这些高表达的GFPuv和aadK基因可以作为这两种生物的遗传标记。(C) 2003欧洲微生物学会联合会。Elsevier Science B.V.版权所有。
A gene expression system for both Bacillus subtilis and Escherichia coli was developed. The expression vector, pHASH102, produces any combination of promoter and open reading frame to be expressed based on the T-extended cloning method. Because the pHASH series vectors are designed to shuttle between the genome and a high copy plasmid in B. subtilis, the expression profiles of copy number dependence can be examined systematically. We demonstrated that vectors with Pr, Pspac, and PS10 promoters are suitable for the overexpression of GFPuv. Moreover, aadK encoding aminoglycoside 6-adenylyltransferase (a streptomycin-resistance gene) of B. subtilis was successfully overexpressed in both B. subtilis and E coli. These highly expressed GFPuv and aadK genes can be used as a genetic marker for both organisms. (C) 2003 Federation of European Microbiological Societies. Published by Elsevier Science B.V. All rights reserved.