FLUORESCENCE PROJECT Homepage

The Fluorescence Project entails mutating gene of interest mRFP1, then expressing the mutant variants in E. coli to determine the new phenotype, if any, after mutation. The Fluorescence Project Overview diagram shows the molecular genetics involved in cloning the mutated genes into a vector and transforming the vector into E. coli bacteria, after which the genes can be expressed to screen for changes to how the bacteria fluoresce, if at all. (Many mutations simply break the fluorescence protein in some way.) We’ll qualitatively assess E. coli colonies for the color and intensity of fluorescence, and quantify the emission spectrum peak wavelength to determine if it changed from the original gene expression. Finally, we’ll sequence the mutant variants of the gene to match any changes to phenotype with changes in the DNA and amino acid sequences. During the semester, each team will work with a single ancestral mRFP1 variant and screen 5  mutated variants.

Readings:

Course Description:

This course is based around a term long project that will explore aspects of molecular genetics, evolution, bioengineering, and heredity using fluorescent proteins in a microbial model system. Through this process students will explore important genetics concepts and implement techniques commonly used to generate new knowledge in the field. We will also explore relevant published literature and practice scientific writing in both lab notebook and lab report formats.

As with all research, we will begin with a question and then follow the scientific method to design and conduct an experiment, analyze the data, and draw a conclusion. Because we’ll be conducting original research, we’ll probably also bump into the primary frustrations of scientific research—assays that require troubleshooting, delays when protocols don’t work perfectly at first pass, and results that don’t match our thinking about the system. We’ll do this because asking real questions in a relevant study system is what scientists do, and learning how to navigate the process and solve the ensuing problems is the best training you can have  for your senior research experience and to pursue careers in scientific research, medicine & human health, or other fields that require problem solving and logic.

Course Goals: By the end of this course, you will be able to:

  • Answer a genetics research question using PCR, gel electrophoresis, sequencing, and other molecular genetics lab techniques.
  • Identify and follow the steps of the scientific method to address a scientific question.
  • Follow, interpret, explain, and (probably) troubleshoot genetics lab protocols and techiniques.
  • Cite relevant genetics primary literature.
  • Write effective and accurate notebook entries, and lab reports in the style accepted by genetics scientific journals.
  • Use appropriate lab safety standards and precautions.