title: pSW2, a Novel Low-Temperature-Inducible Gene Expression Vector Based on a Filamentous Phage of the Deep-Sea Bacterium Shewanella piezotolerans WP3.
abstract: 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.
authors.affiliation: State Key Laboratory of Microbial Metabolism, School of Life Sciences and Biotechnology, Shanghai Jiao Tong University, Shanghai, People's Republic of China College of Life Science, Fujian Normal University, Fuzhou, People's Republic of China.
authors.forename: Xin-Wei
authors.initials: XW
authors.lastname: Yang
authors.affiliation: State Key Laboratory of Microbial Metabolism, School of Life Sciences and Biotechnology, Shanghai Jiao Tong University, Shanghai, People's Republic of China.
authors.forename: Hua-Hua
authors.initials: HH
authors.lastname: Jian
authors.affiliation: State Key Laboratory of Microbial Metabolism, School of Life Sciences and Biotechnology, Shanghai Jiao Tong University, Shanghai, People's Republic of China fengpingw@sjtu.edu.cn.
authors.forename: Feng-Ping
authors.initials: FP
authors.lastname: Wang
mesh_terms: D019677:Artificial Gene Fusion; D004261:DNA Replication; D004926:Escherichia coli / Q000235:genetics*; D015964:Gene Expression Regulation, Bacterial / Q000528:radiation effects*; D017930:Genes, Reporter; D005822:Genetic Vectors; D005827:Genetics, Microbial / Q000379:methods; D049452:Green Fluorescent Proteins / Q000032:analysis / Q000235:genetics; D017912:Inovirus / Q000235:genetics* / Q000302:isolation & purification; D008967:Molecular Biology / Q000379:methods; D010957:Plasmids; D011401:Promoter Regions, Genetic; D011995:Recombination, Genetic; D020592:Shewanella / Q000235:genetics* / Q000821:virology*; D013696:Temperature
chemical_list: D049452:Green Fluorescent Proteins