Swimming through the Yeast Cell

Graduate Fellow Sarah Burnett & Teacher Partner David Buse

Students explore inside the cell, looking at protein-protein interactions.

In this lab and lecture the students will have a better idea of how plasmids can be inserted into the yeast and then can monitor their interaction in the form of proteins by using the yeast two hybrid system. This lab will give students show how DNA (in the form of bacterial plasmids) is incorporated (transformed) into a yeast cell and the then transcribed and translated into a protein product in the form of a reporter (blue color). Students will be able to analyze the protein interactions by the colonies turning blue after just 3 days in an incubator if there is a positive interaction between proteins.

In this activity, students will review the concept of transformation, transcription, translation, and gene regulation by a paper modeling activity.  It is useful to have done the bacterial transformation of fluorescent proteins lab (Sciencebridge) before this activity. They will then do this exercise using the model organism, yeast. The lab activity will be in groups of 2-4 students with minimal lab materials needed on their benches. Students will be assessed through a series of multiple choice questions which can be given before and after the lab and modeling sequence to monitor understanding and learning and can make poster presentations summarizing the interactions

The timing is as follows: optional “Is Yeast Alive?” activity to introduce the model organism (30-1hr); transformation procedure with embedded modeling activity (1-2hrs); third day review of procedures and results (0.5hrs to 1hr).

GRADE LEVEL: 9-12 Biology, AP Biology

DURATION: two to three periods

LEARNING OBJECTIVES:

Students will be able to:

• Be introduced to and operate standard lab equipment vortex, microcentrifuge, shaker and add correct pre-made reagents to tubes in a without cross contamination.
• Identify a change in a gene can lead to change in protein, which then can affect the binding of other proteins to it. Students will have a worksheet demonstrating this.
• Demonstrate knowledge of transcription, translation by laboratory methodology and by paper-based modeling
• Be able to apply knowledge from bacterial transformation to yeast transformation.