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Abstract

As an common pollutant source of industry, low concentration of copper ions can do harm to many creatures. Whereas, Escherichia coli has high tolerance for copper through specific systems. There are copA promoter,marO（operator of mar locus） and cueO promoter existing in E.Coli, all of which are directly or indirectly regulated by copper ions. The most sensitive and effective one (promoter vgb)was chosen to trigger the sequential reactions. Glutathione is an intracellular redox-active tripeptide thiol which can effectively detoxify the metal ions by chelation. It can be synthesized by gshF, an coding sequence derived from Streptococcus thermophilus, which can unify both necessary reactions in one platform.In addition, we also utilize sgRNA-dCas9 system to enhance the efficiency of chelate cupric and cuprous by repressing the expression of copA. With all these design, an automatically and efficiently copper-preventing guard have been created successfully.