Constructed molecular sensor to enhance metal detection by bacterial ribosomal switch–ion channel protein interaction
Molecular biosensors are useful tools that detect metal ions or other potentially toxic chemicals. However, the efficiency of conventional sensors is limited in mixed metals substrates, which is the common way they are found in nature. The use of biosensors constructed from genetically modified living microbial systems has the potential of providing sensitive detection systems for specific toxic targets. Consequently, our investigation was aimed at assembling different genetic building blocks to produce a focused microbial biosensor with the ability to detect specific metals. This objective was achieved by using a synthetic biology approach. Our genetic building blocks, including a synchronized ribosomal switch–iron ion channel, along with sequences of promoters, metal-binding proteins (Fe, Pb), ribosomal binding sites, yellow fluorescence reporter protein (YFRP), and terminators, were constructed within the same biobrick in Escherichia coli. We used an rpoS ribosomal switch containing an aptamer, which responds to the specific metal ligands, in synchronization with an iron ion channel, TonB. This switch significantly stimulates translation, as expressed by higher fluorescence, number of colonies, and concentration of RNA in E. coli. The positive results show the effectiveness of using genetically tailored synchronized ribosomal switch–ion channels to construct microbial biosensors to detect specific metals, as tested in iron solutions.
► Ensembles of different genetic parts for construction of molecular biosensor. ► Use of ribosomal switch and ion channels proteins for enhancing metal detection by molecular biosensor. ► Use of relationship between colony forming unit and ATP as a parameter to confirm the effect of the different interactive genetic parts for detection of metals.
Journal: Journal of Biotechnology - Volume 158, Issues 1–2, 31 March 2012, Pages 1–7