There is a growing consensus that chemistry students benefit from first hand research experiences as opposed to traditional cookbook laboratory experiments. Introduction of research experiences in Biochemistry teaching labs has somewhat lagged behind.  By taking advantage of BioBrick Assembly, students can design simple biosensors that consist of a metal-sensitive promoter and a reporter gene.  As a proof of concept, students were asked to make an Arsenic biosensor.  Arsenic biosensors are of interest to the local community as Arsenic levels in the ground water are typically right at the accepted Environmental Protection Agency levels of 0.01 ppm. Being able to detect microquantities of As could open doors to work with local water purification and chemical plants, as well as benefit the general population of Weatherford, OK.  The use of BioBricks is not a new procedure; however, its use as a teaching tool is fairly new to Biochemistry teaching labs.  There are thousands of potential BioBrick promoter and reporter gene possibilities for this experiment.  The idea is for students to design a sensor based on a set of provided environmental conditions, construct their biosensor, and test the sensitivity of their promoters, yielding endless possibilities.   Arsenic biosensors were developed using synthetic biology techniques.  Several reporter, promoter and destination plasmid BioBricks were obtained.  EcoR1 and Spe1 restriction enzymes were used to hydrolyze the DNA of individual promoter genes, reporter genes, and destination plasmids.  The reporter used, K592009, is a blue chromoprotein that fluoresces blue in the presence of arsenate, indicative of the success of the reaction.  From the restriction enzyme digests, the three reactions were combined with DNA Ligase and incubated. The newly constructed plasmids were transformed into appropriate strains of E. coli. and plated on different arsenic containing media.  In this way, all students learn key biochemical techniques including DNA manipulation with restriction enzymes and ligases, and subsequent analysis of plasmid DNA construction using Agarose gel electrophoresis while contributing to the greater good. In addition to the experiment itself, a portion of the lab discussion is devoted to the exploration of different bioethics topics.  Future directions will include the investigation of the perceptions of societal and environmental impacts of BioBrick assembly by use of a pre/post-experiment survey. <br/><br/>

BioBrick assembly of metal biosensors as a student research experience in biochemistry teaching labs

251st National Meeting (2016)

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