Team:NJAU China/Project
From 2014.igem.org
Overview
Description
For eliminating copper contaminant automatically, the E.coli is supposed to make response as soon as the concentration of copper is excessive in the environment. Sensing and cooperating with copper ion,some special nucleotide sequences on E.coli chromosome can regulate transcription of downstream. Among those copper-sensing sequences, we will choose the most sensitive、effective sequence as a copper sensor to promote sequential reactions.
In our project,we put the concentrations on three candidates:
MarO(THE marRAB OPERON),an regulon which regulates the multiple antibiotic resistance of E.coli, has been reported as a copper sensor to be controlled by MarR.
CopA and CueO ,both of them involves in copper tolerance of E.coli and are activated by CueR in response to excessive concentration of copper.
Our standards and methods for choosing the best one are the sensitiveness and effectiveness.
For the facilitating part, we use vbg promoter to assure that the sequential reactions wouldn’t be actived until the concentration of GSH has accumulated to some point. Therefore, when CopA system are triggered and the concentration of Cu2+ are rise, which has been proved in an article, we can eliminate the copper contaminant with high efficiency. At the same time, we can keep the E.coli safe.
We stabilized the E.coli so that they wouldn’t be released to the environment.