miniPCR® BioBits® Protein Structure and Function

Examine the primary, secondary, and tertiary structures of fluorescent proteins.

• Techniques: Micropipetting, fluorescence visualization, cell-free protein synthesis
• Topics: Levels of protein structure, protein structure and function relationships, fluorescence, protein synthesis, protein analysis
• Time required: 30 minutes during 1 class period, followed by 10 minutes for observation 8-72 hours later
• Level: General high school through college

Proteins are essential tools for life. But how does a simple string of amino acids fold into an intricate three-dimensional structure capable of performing a specific task, such as emitting fluorescence?

Using fluorescent proteins as a model, students will explore a protein’s primary, secondary and tertiary structure and relate those structures to protein function in an integrative, hands-on lab. First, students will compare different fluorescent protein sequences at the primary, secondary, and tertiary levels and use that analysis to predict protein functions. Students will then express these fluorescent proteins using a simple cell-free protocol to test their functional predictions. This activity serves as an excellent interactive introduction to the basics of protein structure and function, as well as a springboard for discussing more advanced topics such as protein engineering.

Required equipment (one per lab group):
- Micropipettes and tips
- P51™ Molecular Fluorescence Viewer or other blue light transilluminator with an orange filter (460 - 480 nm, e.g. blueBox™ or blueGel™)

Storage notes:
- Reagents require freezer storage
- Reagents are stable for 6 months when stored in the freezer
- If a freezer is not available, BioBits® pellets are viable for three months when stored in a refrigerator

 : The kit contains the following reagents for 8 lab groups: 32 BioBits® pellets in 8-tube PCR strips; DNA sample FP1; DNA sample FP2; DNA sample FP3; nuclease-free water; 10 disposable UV flashlights.