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首页> 外文期刊>Applied and Environmental Microbiology >pSW2, a Novel Low-Temperature-Inducible Gene Expression Vector Based on a Filamentous Phage of the Deep-Sea Bacterium Shewanella piezotolerans WP3
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pSW2, a Novel Low-Temperature-Inducible Gene Expression Vector Based on a Filamentous Phage of the Deep-Sea Bacterium Shewanella piezotolerans WP3

机译：pSW2，一种基于深海细菌Shewanella piezotolerans WP3的丝状噬菌体的新型低温诱导基因表达载体

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摘要

A low-temperature-inducible protein expression vector (pSW2) based on a filamentous phage (SW1) of the deep-sea bacterium Shewanella piezotolerans WP3 was constructed. This vector replicated stably in Escherichia coli and Shewanella species, and its copy number increased at low temperatures. The pSW2 vector can be utilized as a complementation plasmid in WP3, and it can also be used for the production of complex cytochromes with multiple heme groups, which has the potential for application for metal ion recovery or bioremediation. Promoters of low-temperature-inducible genes in WP3 were fused into the vector to construct a series of vectors for enhancing protein expression at low temperature. The maximum green fluorescent protein intensity was obtained when the promoter for the hfq gene was used. The WP3/pSW2 system can efficiently produce a patatin-like protein (PLP) from a metagenomic library that tends to form inclusion bodies in E. coli. The yields of PLP in the soluble fraction were 8.3 mg/liter and 4.7 mg/liter of culture at 4°C and 20°C, respectively. Moreover, the pSW2 vector can be broadly utilized in other Shewanella species, such as S. oneidensis and S. psychrophila.
机译：构建了一种基于深海细菌希瓦氏菌WP3的丝状噬菌体（SW1）的低温诱导蛋白表达载体（pSW2）。该载体在大肠杆菌和希瓦氏菌中稳定复制，在低温下其拷贝数增加。 pSW2载体可用作WP3中的互补质粒，也可用于生产具有多个血红素基团的复杂细胞色素，具有用于金属离子回收或生物修复的潜力。将WP3中的低温诱导基因的启动子融合到载体中，以构建一系列用于增强低温下蛋白质表达的载体。当使用hfq基因的启动子时，获得最大的绿色荧光蛋白强度。 WP3 / pSW2系统可以从宏基因组库中高效产生类似patatin的蛋白质（PLP），该库倾向于在大肠杆菌中形成包涵体。在4℃和20℃下，可溶级分中PLP的产量分别为8.3mg /升和4.7mg /升。此外，pSW2载体可广泛用于其他希瓦氏菌属物种中，如one.ensis和S. psychrophila。

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