Using Synthetic Biology in the Detection of Antibiotics

Abstract

Synthetic biology is an innovative field with many applications, including the detection of contaminants of environmental concern. The antibiotic tetracycline is used in cattle and humans, and its presence in the environment may impact development of antibiotic resistance. The objective of our research was to construct and test a tetracycline sensitive bioreporter by combining a tetracycline sensitive promoter and the reporter gene, amiCP, via recombinant DNA technology. The reporter gene amiCP encodes blue chromoprotein and produces a visible color when expressed. We hypothesize that our construct could be used to report the presence of tetracycline via repression of amiCP expression when in the presence of tetracycline. We made a construct expressing amiCP under the regulation of a constitutive promoter as a control. To evaluate our constructs, we exposed the test and control constructs to tetracycline, and detected amiCP expression. The control appeared blue; however, the test construct did not show visible signs of ampCP expression in any condition. Upon further investigation, we discovered that the tetracycline promoter lacked a ribosomal binding site; therefore, although expression of ampCP occurred, production of blue chromoprotein could not occur. Future work is required to add a ribosomal binding site to our construct.