20.109:Module 4

Module 4

Instructors: Angela Belcher and Natalie Kuldell

TA: Chandni Valiathan

In this experimental module we will study an unusual protein, one that allows yeast to bind a metal, gold. Over the next few weeks we will purify yeast based on this binding property, and then we’ll vary some experimental condition to improve the yeast/gold interaction. Using your optimized conditions, we will screen a library of yeast to isolate a new gold-binding strain. The DNA encoding the new gold-binding protein will be sequenced and, with any luck, we’ll elucidate some amino acid requirements for the yeast/metal interaction.

S. cerevisiae shown budding (top) and bound to CdS (bottom)

Module 4 Day 1: Screening library
Module 4 Day 2: Optimizing panning
Day 3: Rescreening gold binders
Day 4: PCR of gold binding candidates
Day 5: Student presentations
Day 6: Sequence analysis