Team:NJAU China

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Abstract

As copper is an essential element for industry, low concentration of it can do harm to many creatures. Whereas, the Escherichia coli has high tolerance for copper through specific systems. There are copA promoter、marO(operator of mar locus)、cueO promoter existed in E.Coli, both are directly or indirectly regulated by copper ions. The most sensitive and effective would be chosen to trigger the sequential reactions. Glutathione is an intracellular redox-active tripeptide thiol which can effectively detoxify the metal ions by chelation. Glutathione can be synthesized by GshF, an enzyme derived from Streptococcus thermophilus, which can unifies 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. Finally, an automatically and efficiently copper-preventing guard are hoped to built.