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Broadening Horizons and Teaching Basic Biology Through Cell-Free Synthesis of Green Fluorescent Protein in a High School Laboratory Course

Abstract

Cell-free protein synthesis (CFPS) has emerged as a practical method for producing a broad variety of proteins. In addition, the direct accessibility to the reaction environment makes CFPS particularly suitable as a learning vehicle for fundamental biological concepts. Here, we describe its implementation as a teaching tool for a high school laboratory course. Ninety students in a biotechnology class used CFPS to study the effects of the concentrations of amino acids, cell extract, DNA, and the energy source on accumulation of active super-folder green fluorescent protein. Students estimated product concentrations simply by comparing solution colors to a printed green color gradient. This simple and inexpensive method allows for immediate measurements, and 26 of the 30 groups observed measurable product concentrations within 60 min. These student-generated data were then discussed to illustrate concepts of data analysis such as outliers and standard deviation. We also combined the laboratory experience with a visit to a university campus that included a laboratory tour and a college-style lecture. Our overall objective was to excite the students about the scientific enterprise and to instill a sense of personal relevance and attainability so that these students could realistically consider technical careers.

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Acknowledgments

We acknowledge the students in the biotechnology class at Sequoia High School for conducting the experiments. We thank Dr. William Yang for the T7 RNA polymerase and for his assistance in preparing the cell extract. Thanks also to Dr. Alyssa Bingham, Dr. Bertrand Lui, and Dr. Phillip Smith for helping with the laboratory tours at Stanford University.

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Correspondence to James R. Swartz.

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Albayrak, C., Jones, K.C. & Swartz, J.R. Broadening Horizons and Teaching Basic Biology Through Cell-Free Synthesis of Green Fluorescent Protein in a High School Laboratory Course. J Sci Educ Technol 22, 963–973 (2013). https://doi.org/10.1007/s10956-013-9442-z

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  • DOI: https://doi.org/10.1007/s10956-013-9442-z

Keywords

  • High school
  • Cell-free protein synthesis (CFPS)
  • PANOx SP
  • Escherichia coli
  • sfGFP
  • Green fluorescent protein
  • Transcription–translation